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  • Articles: DFG German National Licenses  (1,141)
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  • Articles: DFG German National Licenses  (1,141)
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  • 1
    Electronic Resource
    Electronic Resource
    On the ultrastructure of ependymomas — A semiquantitative analysis of diagnostic criteria in 21 cases with special reference to glycogen as a marker (1988)
    Springer
    Neurosurgical review 11 (1988), S. 67-76 
    Details
    ISSN: 1437-2320
    Keywords: Electron microscopy ; ependymoma ; glycogen ; ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We report our results on the ultrastructure of 21 ependymomas and establish the following diagnostic criteria: 1. Glycogen is the most frequently encountered criterion, followed by desmosomes, cilia, basal bodies and microvilli. Fifteen tumors had 3 or more of these features. 2. The allegedly typical nuclear pattern was found in only 6 cases. 3. Special ultrastructural features seen include basement membranes in a mid-thoracic ependymoma. Furthermore we propose the hypothesis that glycogen might be involved in cilia assembly.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01795697
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  • 2
    Electronic Resource
    Electronic Resource
    Origin and direction of replication in mitochondrial plasmid DNAs of broad bean,Vicia faba (1988)
    Springer
    Current genetics 14 (1988), S. 163-170 
    Details
    ISSN: 1432-0983
    Keywords: Plant mtDNA ; Electron microscopy ; Restriction enzymes ; Hairpin structures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Broad bean (Vicia faba) mitochondrial DNA (mtDNA) includes three circular plasmids: mt-plasmid 1 (1,704 ntp), mt-plasmid 2 (1,695 ntp) and mt-plasmid 3 (1,476 ntp). Partially replicated circular forms of these mt-plasmids have been observed in electron microscope preparations. Restriction enzymes that cleave either mt-plasmid 2 (but not mt-plasmids 1 and 3) or mt-plasmid 3 (but not mt-plasmids 1 and 2) were used to generate linear forms of partially replicated mt-plasmid 2 and mt-plasmid 3 molecules. Analyses of these linearized replicative intermediates, observed by electron microscopy, indicated that in both mt-plasmid 2 and mt-plasmid 3 replication originates at a specific location and proceeds in the same, single direction around the molecules. The replication origins of mt-plasmid 2 and mt-plasmid 3 map close to sequences that can fold into hairpin structures.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00569340
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  • 3
    Electronic Resource
    Electronic Resource
    Malignant peripheral neuroectodermal tumours of bone other than Askin's neoplasm: characterization of 14 new cases with immunohistochemistry and electron microscopy (1988)
    Springer
    Virchows Archiv 412 (1988), S. 421-430 
    Details
    ISSN: 1432-2307
    Keywords: Neuroectodermal bone tumours ; Immunochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Peripheral neuroectodermal tumours (PNET) of bone are rare and mimick those seen in soft tissue (peripheral neuroepithelioma of soft tissue). Their differential diagnosis from Ewing's sarcoma (Es) is extremely difficult by optical means. Here we report 14 new cases of PNET of bone (other than Askin's neoplasm) located primarily in the limbs, pelvic girdle and scapula. Clinically and radiologically they displayed Ewing's sarcoma-like features: mean age was 14.4 years, male/ female ratio being 3:11. Metastasis was present in 6 cases at diagnosis (5 with bone metastasis). Prognosis was poor; thirteen patients died; only one with a metatarsal located tumour is alive and free of disease. The mean survival rate was 25 months following diagnosis and treatment with radio- and multimodal chemotherapy. Histologically the 14 cases displayed Homer-Wright rosettes and pseudorosette-like structures, as well as a fibrillary background and lobular pattern. Immunohistochemistry revealed positivity in a number of neural markers when using paraffin-embedded material: NSE, B-2-microglobuline, HNK-1 (leu-7) and E-36 antibodies. At EM level the cell cytoplasms evidenced dense-core granules with neurosecretion, neurotubules and intermediate filaments like those seen in peripheral neuroepithelioma.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00750576
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  • 4
    Electronic Resource
    Electronic Resource
    Alveolar soft part sarcoma (1988)
    Springer
    Virchows Archiv 412 (1988), S. 499-513 
    Details
    ISSN: 1432-2307
    Keywords: Alveolar soft part sarcoma ; Cytology ; Electron microscopy ; Immunohistochemistry ; DNA analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The type, differentiation and histogenesis of the tumor cells of alveolar soft part sarcoma (ASPS) have been analyzed in a series of ten cases by a light-microscopic, ultrastructural, immunohistochemical and cytologic investigation and quantitative DNA analysis. Four tumors deviated from ordinary ASPS: three were wholly or partly of the so-called pleomorphic variant of ASPS and a fourth tumor showed calcifications of the psammoma body type. The ultrastructural findings and immunohistochemical demonstration of desmin supported the hypothesis of a rhabdomyomatous differentiation and gave no support to epithelial (negative immunoreactions for cytokeratins, epithelial membrane antigen, HMFG-1 and -2, tissue polypeptide antigen (TPA)) or neuroectodermal (negative for S-100 protein, glial fibrillary acidic protein, neurofilaments) differentiation. The negative immunoreactions for vimentin and myoglobin and the positive reaction for neuron specific enolase (NSE) do not exclude a rhabdomyomatous differentiation since in rhabdomyosarcomas the undifferentiated rhabdomyoblasts generally contain vimentin and the differentiated tumor cells contain myoglobin and rhabdomyosarcoma has previously been reported as being positive for NSE. The production of external lamina material peripherally in the tumor cell nests and around vessels in the vascular septa was demonstrated both ultrastructurally and by immunohistochemistry using antibodies against collagen IV and laminin. The cytologic appearance in smears obtained by fine-needle aspiration from a case of the pleomorphic variant showed some resemblance to that of a carcinoma. The seven tumors with an ordinary cell appearance were found to show a diploid DNA-distribution at a quantitative analysis performed on paraffin sections, while the three tumors wholly or partly of the pleomorphic type showed an additional tetraploid peak.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00844286
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  • 5
    Electronic Resource
    Electronic Resource
    Ultrastructural and immunohistochemical findings in oral hairy leukoplakia (1988)
    Springer
    Virchows Archiv 412 (1988), S. 533-542 
    Details
    ISSN: 1432-2307
    Keywords: Hairy leukoplakia ; Electron microscopy ; Epstein-Barr virus ; Candida albicans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Three cases of HL from the lateral border of the tongue of male homosexual AIDS patients were investigated by thin section electron microscopy. Keratinocytes contained condensed chromatin in their pyknotic nuclei and a few organelles in the oedematous cytoplasm. Chromatin was in close association to the nuclear membrane and showed a punched-out appearance. Particles typical of the herpes virus group were abundant in the upper two thirds of the epithelium in all three cases. Virus particles were seen frequently in the nuclei of the ballooned keratinocytes, but rarely in cells containing Candida albicans. Viral nucleocapsids were observed budding at the inner nuclear membrane, thereby acquiring the prospective viral envelope. Complete, enveloped virions were found in the endoplasmic reticulum and in the extracellular space. These virions were identified immunohistochemically as Epstein-Barr virus (EBV) using two monoclonal antibodies directed against EBV capsid and membrane antigen, respectively. Candida albicans was observed in the stratum corneum and in the upper layer of the stratum spinosum. Special cytoplasmic tubular structures arranged in parallel bundles were found in koilocytotic cells in addition to characteristic membrane structures composed of undulating convoluted membranes. Epithelial basement membranes were always intact.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00844289
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  • 6
    Electronic Resource
    Electronic Resource
    Ultrastructural features of acute monoblastic leukaemia cells: A multivariate morphometric analysis (1988)
    Springer
    Virchows Archiv 414 (1988), S. 21-27 
    Details
    ISSN: 1432-2307
    Keywords: Monoblastic leukaemia ; Electron microscopy ; Morphometry ; Multivariate statistics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Morphometric studies were carried out on five ultrastructural measures of size or quantity of some of the intracellular organelles in monoblasts obtained from six patients diagnosed as having acute monoblastic leukaemia and also on monocytes from six normal controls. The morphometric measures were analysed using a one way multivariate analysis of variance (MANOVA) to see whether acute monoblastic leukaemic cells differed from those of normals. It was found that there was a highly significant decrease both in the surface to volume ratio of mitochondria and also in the surface to volume ratio of the nucleus. The possible physiological significance of these structural changes is stressed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00749734
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  • 7
    Electronic Resource
    Electronic Resource
    Osteosarcoma with a clear-cell component (1988)
    Springer
    Virchows Archiv 412 (1988), S. 273-279 
    Details
    ISSN: 1432-2307
    Keywords: Osteosarcoma ; Clear cell ; Histopathology ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The reports concerns the light microscopical and ultrastructural findings obtained in three conventional osteosarcomas with an unusually high admixture of clear cells, whose presence appeared to be responsible for the marked change in the histological pattern of these tumours. In the tumours with a prevailing fibroblastic component the clear cells were either irregularly scattered throughout the tumour in the form of small groups, or they formed large groups sharply demarcated against the fusicellular areas of the tumours. In two cases it was shown that their cytoplasm contained exaggerated glycogen deposits accompanied by the formation of glycogen-containing phagolysosomes and occasional empty vacuoles. In the third case the clear cells showed vacuolar degeneration with numerous single-membrane-bound, empty vacuoles. In contrast to the clear-cell chondrosarcoma we did not find S-100 protein in clear cells of our osteosarcomas. Such findings could be particularly significant in the differential diagnosis of bone tumours.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00737152
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  • 8
    Electronic Resource
    Electronic Resource
    Oncocytomas and null cell adenomas of the human pituitary: Morphometric and in vitro functional comparison (1988)
    Springer
    Virchows Archiv 413 (1988), S. 333-339 
    Details
    ISSN: 1432-2307
    Keywords: Electron microscopy ; Morphometry ; Null cell adenoma ; Oncocytoma ; Pituitary adenoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In this study, light microscopic and ultrastructural morphometric features of oncocytomas and null cell adenomas were compared and the morphometric data were correlated with in vitro endocrine activity. All tumours were unassociated with clinical or biochemical evidence of hormone excess and were diagnosed as oncocytomas or null cell adenomas, using histology, immunohistochemistry and electron microscopy. In oncocytomas, when compared with null cell adenomas, light microscopic morphometry revealed that total cell areas were significantly larger and nuclear cytoplasmic ratios were smaller due to an increase in cytoplasmic areas. Ultrastructural morphometry disclosed an abundance of mitochondria in oncocytomas. Absolute volumes of cytoplasmic organelles per cell were not reduced in oncocytomas compared with those of null cell adenomas. These results indicate that accumulating mitochondria do not replace other cytoplasmic organelles, and furthermore that the functional potential of oncocytomas is not lost. In vitro study demonstrated the production of pituitary hormones, primarily gonadotropins in oncocytomas and null cell adenomas. It can be concluded that oncocytomas, which represent the final stage of oncocytic transformation, have a close relationship with null cell adenomas based on morphometric comparison as well as in vitro studies.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00783026
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  • 9
    Electronic Resource
    Electronic Resource
    Small cell neuroendocrine carcinoma of the urinary bladder (1988)
    Springer
    Virchows Archiv 413 (1988), S. 505-512 
    Details
    ISSN: 1432-2307
    Keywords: Neuroendocrine differentiation ; Immunohistochemistry ; Electron microscopy ; Small cell undifferentiated bladder carcinoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Small cell carcinoma with the histological appearance of pulmonary small cell carcinoma is a rare tumour in the urinary bladder. In previous case reports the neuroendocrine nature of small cell bladder carcinoma has been accepted, but on review the evidence for true neuroendocrine differentiation appears unsatisfactory. In this study the histological, immunohistochemical and ultrastructural characteristics of three cases of small cell carcinoma of the urinary bladder are described. Ultrastructurally, the cytoplasm of all three tumours contained neurosecretory-type granules and each of the tumours demonstrated positive immunoreaction for two or more neuroendocrine markers, from a panel including neuron-specific enolase, chromogranin A, Leu-7, bombesin and synaptophysin. Although the combination of ultrastructural and immunohistochemical examination obviously offers the strongest evidence in establishing neuroendocrine differentiation, it is argued that immunohistochemistry alone may also yield important information in demonstrating a neuroendocrine nature, provided that at least neuron-specific enolase and synaptophysin are included as markers. The clinical relevance of identifying neuroendocrine differentiation in small cell bladder carcinoma is suggested by the favourable response to combination chemotherapy in two of our cases.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00750391
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  • 10
    Electronic Resource
    Electronic Resource
    Do folliculo-stellate adenomas of the pituitary gland exist or are they intrasellar meningiomas? (1988)
    Springer
    Acta neuropathologica 77 (1988), S. 219-223 
    Details
    ISSN: 1432-0533
    Keywords: Meningioma ; Folliculo-stellate cell ; Pituitary neoplasm ; Electron microscopy ; Immunochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A tumor arising in the pituitary fossa and having some of the histological and ultrastructural features of a recently described tumor, purportedly originating from the folliculo-stellate cells of the anterior pituitary, is presented. The results of our ultrastructural and immunohistochemical studies, however, favored a meningeal origin and suggested that the neoplasm was most likely a secretory meningioma.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00687435
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  • 11
    Electronic Resource
    Electronic Resource
    Structural changes in the rat brain after carotid infusions of hyperosmolar solutions (1988)
    Springer
    Acta neuropathologica 77 (1988), S. 5-13 
    Details
    ISSN: 1432-0533
    Keywords: Blood-brain barrier ; Hyperosmolar solutions ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Infusion of hypertonic solutions into the carotid artery is one method by which the blood-brain barrier (BBB) can be opened transiently in experimental animals. This technique has also been tried in clinical situations in which an enhanced uptake of intravenously injected chemotherapeutic drugs into the brain is desired. We have previously found that infusion of hypertonic mannitol or urea into the carotid artery of the rat, leading to a BBB opening, is associated with light microscopic signs of cellular damage in the brain parenchyma. An electron microscopic study has now been made to obtain more detailed information about the events taking place in the rat brain 1 to 72 h after an intracarotid infusion of hyperosmolar solution of mannitol. Toluidine blue-stained semithin epon sections were also available for high-resolution light microscopy of brain samples from urea-infused animals. Intravenously injected Evan's blue dye was used to confirm that BBB opening had occurred as a consequence of the carotid infusions. The infused hemispheres had numerous structural changes. The dominating light microscopic alteration was the presence of multifocal lesions in the gray or the white matter with closely packed microvacuoles causing status spongiosus. Ultrastructurally the microvacuoles corresponded to very pronounced watery swelling of astrocytic processes and to a minor degree to expansion of dendrites and axons. There was also a light or moderate perivascular astrocytic swelling. In the “spongy” lesions as well as occasionally in non-vacuolated parts of the cerebral cortex, there were collapsed electron-dense neurons with pronounced mitochondrial alterations such as severe swelling associated with rupture of christae. Rats with a survival period of 24 h or 72 h showed several disintegrating neurons and accumulation of macrophages. This study shows that carotid infusion of hypertonic mannitol in the rat may cause pronounced neuronal changes as well as multifocal astrocytic swelling. The severity of the nerve cell changes and the presence of macrophages indicate that some of the alterations are irreversible and thus, such a procedure may not be as safe as previously suggested.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00688236
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  • 12
    Electronic Resource
    Electronic Resource
    Nerve cell injury in the brain of stroke-prone spontaneously hypertensive rats (1988)
    Springer
    Acta neuropathologica 76 (1988), S. 227-237 
    Details
    ISSN: 1432-0533
    Keywords: Stroke-prone spontaneously hypertensive rats ; Blood-brain barrier ; Brain edema ; Nerve cell injury ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The brain lesions in stroke-prone spontaneously hypertensive rats (SHRSP) are characterized by multifocal microvascular and spongy-cystic parenchymal alterations particularly in the gray matter. An essential feature of the lesions is the presence of edema with massive extravasation of plasma constituents as evidenced by specific gravity measurements, Evans blue technique and immunohistochemistry. The nerve cell injury occurring in the brain lesions in SHRSP is further characterized by light and electron microscopy in the present study. Two types of neuronal changes were seen within the blood-brain barrier (BBB) leakage sites. A small number of neurons with dark condensed nucleus and cytoplasm were found most often at the periphery of recent lesions. The majority of injured neurons were pale and showed intracellular edema confined to the dendrites and perikarya sparing axons and synapses. Their nuclei were weli preserved with finely dispersed chromatin. The swollen and watery cell processes of neurons and astrocytes gave a spongy appearance to the neuropil. The intracellular edema seemed to result in cytolysis. The results suggest that primary anoxiaischemia is not the major pathogenetic mechanism behind the nerve cell injury in severely hypertensive SHRSP, rather it is the massive BBB leakage and consequent brain edema that causes cytolytic destruction of neurons. Secondary focal ischemia as a consequence of occlusion in microvessels may, however, contribute to the nerve cell destruction.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00687769
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  • 13
    Electronic Resource
    Electronic Resource
    The ultrastructural organization of the isthmic nucleus in Xenopus (1988)
    Springer
    Anatomy and embryology 177 (1988), S. 325-330 
    Details
    ISSN: 1432-0568
    Keywords: Isthmic nucleus ; Electron microscopy ; Tectal ablation ; Xenopus ; Morphometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The isthmic nucleus (IN) of the frog brain forms a linkage, relaying visual information from one tectum to the other. It receives afferent input from the tectum of the same side and projects bilaterally to both tecta. The ultrastructural features of the tecto-isthmic synaptic connections were studied in young postmetamorphic Xenopus frogs. Most synaptic profiles in the isthmic nucleus have spheroidal vesicles and an asymmetric zone of apposition. Frequently, synaptic glomeruli consisting of up to 8 terminal boutons surrounding a shaft dendrite were observed. The syanptic density in the rostral IN was slightly higher than in the middle or caudal portions. Partial deafferentation by transection of the tectoisthmic pathway or total deafferentation by removal of the tectum was followed by a widespread degeneration of terminals in the ipsilateral IN. In the former case, the density of synapses in the IN decreased initially by about 64%, and then increased by 30 days after operation to about 50% of the normal synaptic density. After tectal removal, all the terminal boutons in the isthmic neuropil degenerated by 3 days after operation. These studies, along with recent findings, indicate that most, if not all, of the afferent fibres to IN are of tectal origin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00315840
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  • 14
    Electronic Resource
    Electronic Resource
    Fine structural survey of Gudden's tegmental nuclei in the rat: cytology and axosomatic synapses (1988)
    Springer
    Anatomy and embryology 177 (1988), S. 485-493 
    Details
    ISSN: 1432-0568
    Keywords: Ventral tegmental nucleus of Gudden ; Dorsal tegmental nucleus of Gudden ; Quantitative morphometric study ; Synaptic organization ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The fine structure of neuronal somata and axosomatic synapses in each subnucleus of Gudden's tegmental nuclei was studied by use of electron microscopy. The pars principalis of the ventral tegmental nucleus of Gudden (TVP) is composed of oblong or triangular, medium sized neurons (11.8x22.6 μm, 211.4 μm2) containing many mitochondria, lysosomes, Golgi apparatus, and rough endoplasmic reticulum composing Nissl bodies. The light oval nucleus with a prominent nucleolus is centrally situated, and indentations of its nuclear envelope are recognized in all neurons. The neuron in the pars ventralis of the dorsal tegmental nucleus of Gudden (TDV) is similar to that in the TVP, but its average size is significantly smaller (10.0x18.8 μm, 151.4 μm2), and its organelles are also less well developed. The pars dorsalis of the dorsal tegmental nucleus of Gudden (TDD) is composed of spindle shaped, small neurons (6.9x16.2 μm, 85.1 μm2) characterized by their irregular shaped nucleus with its invaginated envelope. These neurons have a thin rim of cytoplasm, poorly developed organelles and no Nissl bodies. The average number of axosomatic terminals in a sectional plane is 9.9 in the TVP, 9.6 in the TDV and 2.6 in the TDD, and the bouton covering ratio is 24.3% in the TVP, 26.5% in the TDV and 7.4% in the TDD. The respective percentages of round, flat and pleomorphic type axosomatic terminals were estimated, and the flat type terminals were found to be dominant in the TVP, the pleomorphic type terminals in the TDV, and the round type terminals in the TDD.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00305136
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  • 15
    Electronic Resource
    Electronic Resource
    Quantitative electron microscopic observations on the non-neuronal cells and lipid droplets in the posterior funiculus of the kitten after dorsal rhizotomy (1988)
    Springer
    Anatomy and embryology 178 (1988), S. 95-105 
    Details
    ISSN: 1432-0568
    Keywords: Electron microscopy ; Glial cells ; Perivascular cells ; Lipid droplets ; Wallerian degeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Kittens were subjected to lumbosacral dorsal rhizotomies at the age of 6–8 days postnatally. After postoperative survival times of 1–25 days the number of non-neuronal cells and lipid droplets in each cell type in the posterior funiculus at L1 were counted at the ultrastructural level. Intact control animals were analyzed in the same way. The number of astrocytes and oligodendrocytes decreased with increasing postoperative survival time in the degenerating zone. This was also the case in the white matter of control animals with increasing age of sacrifice. However, in the degenerating zone of operated animals the decrease was more extensive for oligodendrocytes starting at 5 days after surgery, and possibly also for astrocytes at 25 days postoperatively. The number of microglial cells in the degenerating zone was markedly increased 2–10 days after surgery compared to the controls. The number of non-pericytic perivascular cells seemed to be somewhat increased from 9 days after surgery, while the number of pericytes remained unchanged during the experimental period. Lipid droplets in the degenerating white matter were mainly located in microglial cells and astrocytes and only to a small extent in nonpericytic perivascular cells. These findings suggest that lipid material produced during anterograde fiber degeneration in the immature white matter is mainly metabolized in glial cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02463643
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  • 16
    Electronic Resource
    Electronic Resource
    Ultrastructure of preimplantation rabbit embryos exposed to visible light and room temperature (1988)
    Springer
    Anatomy and embryology 178 (1988), S. 229-241 
    Details
    ISSN: 1432-0568
    Keywords: Room temperature ; Visible light ; Preimplantation embryos ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Early cleavage stage embryos (day 1 p.c.) and morulae (day 3 p.c.) of rabbits were exposed to visible (standard) lighting (1600 lx) and room (standard) temperature (23°C) during a 24 h in-vitro culture. Control embryos were cultured in darkness at 37°C. Development was assessed by light and electron microscopy as well as by the cytochemical demonstration of glycogen. In day 1 and day 3 embryos standard temperature induced swelling of the SER and Golgi complex vesicles. Major changes in day 1 embryos consisted of smallish microtubules-like crystalloids, and in day 3 embryos of unusually large SER vesicles. In both embryonic ages cleavage rate and development was more retarded by standard temperature than by standard lighting. Standard lighting, however, led to distinct signs of degeneration and cell death. The mode of cell damage seemed to be different in light exposed early cleavage stages and morulae: In day 1 embryos cytoplasmic degeneration was predominant while the majority of cells in day 3 embryos died by apoptosis. Despite clear indications of cell damage, cleavage rate was not notably impaired compared with non-exposed controls. Glycogen increased during development from cleavage stages to early blastocysts. The distribution was not changed either by exposure to standard temperature nor by standard lighting. The results demonstrate that day 1 embryos were clearly more susceptible to lighting whereas day 3 embryos were more affected by temperature. The mode of damage exerted by both the physical environmental factors was different. Reduction to standard temperature interfered mainly with the organization of the cytoskeleton and intracellular transport of organelles, while exposure to standard lighting led to cell degeneration and death.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00318226
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  • 17
    Electronic Resource
    Electronic Resource
    Heterogeneities of the islets in the rabbit pancreas and the problem of “paracrine” regulation of islet cells (1988)
    Springer
    Anatomy and embryology 178 (1988), S. 297-307 
    Details
    ISSN: 1432-0568
    Keywords: Rabbits ; Islands of Langerhans ; Electron microscopy ; Immunohistochemistry ; Regulation of islands
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In addition to “external” signals conveyed by the circulation or the nervous system, the pancreatic islets obviously are regulated also by “internal” (intra-islet) signals, e.g. by the islet hormones: insulin (B-), glucagon (A-), and somatostatin (D-) cells are able to affect the secretion of the heterologous cell types. It is, however, unclear whether this functional cooperation between islet cells occurs by an intercellular route (paracrinia sensu strictore), by intraislet “portal” vessels, or by the systemic circulation. These likely interactions are limited by islet anatomy. To identify the anatomical basis for the mutual functional relationships between the islet cells, islets of Langerhans in the rabbit pancreas were completely analyzed in immunostained serial semithin (0.5 μm) sections. The islets were found to be largely heterogenous. They were classified in three basic types: a) polycellular islets, composed of all established endocrine cells, and including two subtypes of islets, b) bicellular islets, containing only B- and A-cells or B- and D-cells, and c) monocellular islets, exclusively made up of B-cells. Concerning the modes of paracrine regulation of islet cells, the findings suggest primarely an endocrinous route of transport of the islet peptides to heterologous endocrine cells. The corresponding functional cooperation between islet cells probably is mediated rather by the systemic circulation than by intra-islet portal vessels.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00698661
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  • 18
    Electronic Resource
    Electronic Resource
    Light and electron microscopic investigation of the rat placenta after cadmium administration during pregnancy (1988)
    Springer
    Anatomy and embryology 178 (1988), S. 345-351 
    Details
    ISSN: 1432-0568
    Keywords: Cadmium, Placenta (rat) ; Morphometry ; Light microscopy ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The morphology of the rat placenta was studied after exposure to cadmium chloride during pregnancy, using optimal fixation conditions. In contrast to previous observations, no differences were observed after cadmium administration in relative volume densities of trophoblastic tissue, maternal lacunae, fetal capillaries and connective tissue, nor in trophoblastic thickness or other morphometric features. At the ultrastructural level, the amount of glycogen in trophoblast layer II was elevated in cadmium exposed rats, but other electron microscopic features (amount and localization of lipid, degenerative vesicles, thickness and general appearance of the trophoblastic and endothelial layers and thickening or multiplication of the basal lamina) were not changed. Results obtained from the present experiments do not support the suggestion that cadmium is responsible for structural changes in the placentae of human smokers.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00698665
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  • 19
    Electronic Resource
    Electronic Resource
    Quantitative assessment of taurine-like immunoreactivity in different cell types and processes in rat cerebellum: an electronmicroscopic study based on a postembedding immunogold labelling procedure (1988)
    Springer
    Anatomy and embryology 178 (1988), S. 407-421 
    Details
    ISSN: 1432-0568
    Keywords: Taurine ; Immunocytochemistry ; Quantification ; Electron microscopy ; Cerebellum ; Purkinje cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Ultrathin sections of plastic-embedded rat cerebella were incubated with an antiserum against conjugated taurine and subsequently treated with a secondary antibody coupled to colloidal gold. The density of gold particles in various cellular profiles was calculated with the assistance of a computer. In the cerebellar cortex the highest density was found in the somata, dendrites, and dendritic spines of the Purkinje cells, supporting parallel light-microscopical observations in postembedding stained semithin sections from the same tissue blocks. The remaining profiles could be divided into three groups according to their immunolabelling intensity, in descending order: 1) somata and processes of granule and Golgi cells; 2) somata and processes of stellate, basket, and glial cells, and 3) mossy fiber terminals. In a representative experiment, the structures in the first and second groups showed gold particle densities in the range of 19–25%, and 4–11%, respectively, of that in the Purkinje cell somata (values corrected for background) whereas the particle density in the mossy fiber terminals was not significantly above background level. In the cerebellar nuclei, taurine-like immunoreactivity was concentrated in terminals that typically established symmetric or intermediate type contacts with weakly labelled dendrites and cell bodies. These terminals, which shared the ultrastructural features of Purkinje cell terminals, showed an average gold particle density that was about 60% higher than that of the Purkinje somata. For specificity control, ultrathin sections containing a series of different amino acid conjugates were incubated in the same drops of sera as the tissue sections. The highly selective labelling of the taurine conjugate indicated that the distribution of gold particles in the tissue was not confounded by crossreactivity with GABA, glutamate or other common amino acids but adequately reflected the distribution of fixed taurine. For additional control of specificity, the taurine antiserum was applied to the soluble fraction of a rat brain extract separated by thin layer chromatography. In this system the taurine antiserum stained a single spot that comigrated with free taurine. The present results suggest that all cell types and processes in the rat cerebellum (with the exception of the mossy fiber terminals) contain taurine. However, the concentration of taurine appears to vary considerably among the different cell types and may also differ between different parts of the same neuron.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00306047
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  • 20
    Electronic Resource
    Electronic Resource
    Proliferation of rat intraspinal Schwann cells following tellurium intoxication (1988)
    Springer
    Acta neuropathologica 76 (1988), S. 624-627 
    Details
    ISSN: 1432-0533
    Keywords: X-Irradiation ; Intraspinal Schwann cell ; Myelin ; Autoradiography ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A stable population of intraspinal Schwann cells, which developed follwing early postnatal irradiation of the spinal cord, was challenged by the addition of tellurium (Te) to the diet beginning at 30 days of age. Schwann cells incorporating [3H]thymidine were identified by 1 μm autoradiographs and by conventional electron microscopy of adjacent thin sections. Autoradiographs of areas with Schwann cell myclination showed extensive labelling of cells in the Te-fed animals. In contrast, control animals which were not fed Te showed little evidence of labelled Schwann cells. These data indicate that Schwann cells in the intraspinal environment show a proliferative response to the presence of Te in the rat's diet, as do Schwann cells in their normal extraspinal milieu.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00689602
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  • 21
    Electronic Resource
    Electronic Resource
    Abnormal amianthoid collagen fibers in meningioma (1988)
    Springer
    Acta neuropathologica 76 (1988), S. 638-639 
    Details
    ISSN: 1432-0533
    Keywords: Meningioma ; Electron microscopy ; Collagen ; Amianthoid fibers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Electron microscopic examination of a spinal meningioma demonstrated broad zones with large amianthoid collagen fibers in the tumor. Normal native collagen fibrils were found in small scattered clumps particularly around the blood vessels in the tumor. Such abnormal large collagen fibrils have not been seen previously in meningiomas. The significance of the amianthoid fibers is unknown. The only other unusual feature of the tumor was that it had many pale-staining cells by light microscopy and was classified as a clear cell variant of meningioma.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00689605
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  • 22
    Electronic Resource
    Electronic Resource
    Morphological study of peripheral nerve changes induced by chloroquine treatment (1988)
    Springer
    Acta neuropathologica 75 (1988), S. 253-260 
    Details
    ISSN: 1432-0533
    Keywords: Chloroquine ; Peripheral neuropathy ; Nerve biopsy ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Nerve biopsies were performed in four patients with suspected chloroquine induced neuromyopathy. Three of the patients were treated with high doses of chloroquine for connective tissue disease, while one patient was taking this drug as malaria prophylaxis. Morphological studies demonstrated the presence of segmental demyelination and remyelination in all cases. Cytoplasmic inclusions were observed in Schwann cells, in perineurial and endothelial cells, and in some interstitial cells. They were never observed within axons. Occasional curvilinear profiles were seen in perineurial and Schwann cells. Perineurial calcifications were observed in two cases. The results of this morphological study suggest that chloroquine neuropathy is essentially due primary involvement of Schwann cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00690533
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  • 23
    Electronic Resource
    Electronic Resource
    Human immunodeficiency virus (HIV)-related chronic relapsing inflammatory demyelinating polyneuropathy with multifocal unusual onion bulbs in sural nerve biopsy (1988)
    Springer
    Acta neuropathologica 75 (1988), S. 529-534 
    Details
    ISSN: 1432-0533
    Keywords: HIV-related relapsing polyneuropathy ; Intrathecal HIV antibodies ; Nerve biopsy ; Electron microscopy ; Schwann cell morphology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary This is obviously the first report on a case with a spontaneous sensu strictu relapsing variant of human immunodeficiency virus (HIV)-related polyneuropathy. Its manifestation preceded LAS. Intrathecal HIV-antibodies developed between the most severe third and fourth episode. Analysis of sural nerve biopsy was consistent with a multifocally accentuated chronic inflammatory demyelination, characterized by unusual onion bulb-like Schwann cell formations with irregular voluminous layers, electron density, aggregation of filaments, multiple indented nuclei, and numerous enclosed collagen pockets. A direct or immune-mediated indirect specific influence on Schwann cell morphology by HIV might be discussed. Virus-like particles and ultrastructural markers of HIV were not detectable.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00687143
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  • 24
    Electronic Resource
    Electronic Resource
    A novel intradural extramedullary spinal cord tumor in young dogs (1988)
    Springer
    Acta neuropathologica 75 (1988), S. 402-410 
    Details
    ISSN: 1432-0533
    Keywords: Spinal cord tumor ; Dog ; Electron microscopy ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We have studied an unusual, spontaneous, intradural extramedullary spinal cord tumor in 12 dogs. Animals presented with paraparesis and ataxia early in life (11/12 ranged from 6 to 38 months of age) suggesting that these tumors may be congenital. Various breeds of dogs were represented with four cases in German Shepherds and three in retrievers; there was no sex predisposition. Post-mortem examinations revealed a single intradural mass consistently located between T10 and L2, which produced extensive compression of the spinal cord. Metastasis was never observed and significant pathological changes in other organs were lacking. Microscopic examination revealed solid sheets of ovoid to fusiform cells interspersed with areas of acinar and tubular differentiation. Some areas were rarified and focal squamous metaplasia was observed. Ultrastructural features included the presence of a continuous basal lamina, junctional complexes, microvilli and occasional cilia at the apices of acinar complexes. Immunocytochemical studies did not support a neurectodermal origin. At least 13 case reports of this entity have been previously published and have been designated ependymomas, medulloepitheliomas and neuroepitheliomas. A recent case was diagnosed as a nephroblastoma and we feel that this is an interesting and provocative diagnosis. These tumors could result from remnants of renal primordium which becomes trapped between the dura and the developing spinal cord. However, firm evidence of such a histogenesis is not yet at hand.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00687794
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  • 25
    Electronic Resource
    Electronic Resource
    Ultrastructural analysis of neurofibrillary tangles of Alzheimer's disease using computerized digital processing (1988)
    Springer
    Acta neuropathologica 75 (1988), S. 519-522 
    Details
    ISSN: 1432-0533
    Keywords: Alzheimer's disease ; Neurofibrillary tangle ; Paired helical filament (PHF) ; Electron microscopy ; Computerized digital processing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The ultrastructure of neurofibrillary tangles of Zlzheimer's disease was analyzed by computerized digital processing of electron micrographs. Processing of the electron micrographs consists of four steps: digitizing the electron micrograph, Fourier transformation, noise filtering and inverse Fourier transformation and Laplacian operation. In the present study, we have confirmed that neurofibrillary tangles are composed of a pair of helical filaments (PHF), which appear characteristically as an unbranched rigid structure. The periodicity of PHF is 78nm on the diffractogram. The dimensions of PHF obtained by our analysis, although basically similar to those described earlier by other investigators using conventional techniques, more precisely defines its structural conformation. We have also demonstrated that the spatial relationship of two filaments appears symmetrical after two-way tilting of the specimen about the axis of rotation. Our observations emphasize the importance of digital image processing as an effective tool for structural analytical research in biology and medicine.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00687141
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  • 26
    Electronic Resource
    Electronic Resource
    Ultrastructural pathology of dermal axons and Schwann cells in lysosomal diseases (1988)
    Springer
    Acta neuropathologica 76 (1988), S. 489-495 
    Details
    ISSN: 1432-0533
    Keywords: Dermal nerves ; Electron microscopy ; Lysosomal diseases ; Terminal axons
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Skin tissue specimens, obtained from 60 patients afflicted with a diverse range of lysosomal disorders revealed two groups of lesions within dermal axons, largely unmyelinated ones, particularly within axonal terminals: (1) non-specific mitochondria and dense bodies often enlarging the axonal terminal; and (2) disease-specific lysosomal residual bodies, the latter less frequent depending on the incidence and type of lysosomal disorders, i.e., largely only seen in GM2-gangliosidosis due to hexosaminidase A deficiency and mucolipidosis IV, while the spectrum of lysosomal residual bodies in Schwann cells appeared more variegated, especially due to the occurrence of vacuolar lysosomal residual bodies which were never seen within axons. The most frequent location of abnormal intraaxonal constituents in terminal axons indicates a functionally and morphologically impaired retrograde axonal transport but provides no further evidence as to whether the respective parent nerve cell body has also accumulated lysosomal residual bodies. When studying biopsied skin specimens for diagnosis, axonal terminals beneath the epidermis, about sweat glands, and among smooth muscle cells, ought to be incorporated into a comprehensive electron microscopic examination.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00686388
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  • 27
    Electronic Resource
    Electronic Resource
    The postsynaptic targets of substance P-immunoreactive terminals in the rat neostriatum with particular reference to identified spiny striatonigral neurons (1988)
    Springer
    Experimental brain research 70 (1988), S. 361-377 
    Details
    ISSN: 1432-1106
    Keywords: Neostriatum ; Substance P ; Immunocytochemistry ; Golgi-impregnation ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Substance P-immunoreactive boutons were examined in the electron microscope in sections of the rat neostriatum that contained retrogradely labelled striatonigral neurons and/or Golgi-impregnated medium-size densely spiny neurons. The postsynaptic targets of the immunoreactive boutons were characterized on the basis of ultrastructural features, their projection to the substantia nigra and/or their somato-dendritic morphology. Substance P-immunoreactive axonal boutons formed symmetrical synaptic specializations. Of a total of 233 randomly identified synaptic boutons 72.5% made contact with dendritic shafts, 15% with dendritic spines and 10.7% with perikarya. The ultrastructural characteristics of some of the postsynaptic neuronal perikarya were consistent with their identification as striatal interneurons. Similarly, the observation of some of the substance P-containing terminals in contact with spines, spine-bearing dendritic shafts and perikarya with the ultrastructural characteristics of medium-size densely spiny neurons suggested that one of the targets of substance P-positive terminals are striatal projection neurons. Direct evidence for this was obtained in sections from rats that had received injections of horseradish peroxidase conjugated with wheatgerm agglutinin in the substantia nigra. The perikarya of retrogradely labeled striatonigral neurons were found to receive symmetrical synaptic input from substance P-positive boutons. Ultrastructural analysis of Golgi-impregnated medium-size densely spiny neurons, some of which were also retrogradely labeled from the substantia nigra, demonstrated directly that this class of neuron was postsynaptic to the substance P-immunoreactive boutons. The combination of Golgi-impregnation with substance P-immunocytochemistry made it possible to study the pattern or topography of the substance P-positive input to medium size densely spiny neurons. The substance P-containing boutons made contact predominantly with perikarya and dendritic shafts. This pattern of input is markedly different from that of other identified inputs to medium-size densely spiny neurons.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00248361
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  • 28
    Electronic Resource
    Electronic Resource
    The cholinergic innervation of the rat substantia nigra: a light and electron microscopic immunohistochemical study (1988)
    Springer
    Experimental brain research 72 (1988), S. 178-184 
    Details
    ISSN: 1432-1106
    Keywords: Choline acetyltransferase ; Immunocytochemistry ; Substantia nigra ; Pedunculopontine tegmental nucleus ; Acetylcholine ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Putative cholinergic axons and synaptic endings were demonstrated in the substantia nigra (SN) of the rat by light and electron microscopy on the basis of the localization of choline acetyltransferase (ChAT) immunoreactivity. The distribution of ChAT immunoreactivity in the SN as demonstrated by light microscopy revealed a modest network of ChAT-immunoreactive beaded axons in the SNc, in comparison to a relatively sparse distribution in the SNr. These axonal profiles were most dense in the middle of the rostral-caudal extent of the SNc and appeared to be concentrated in the middle third of the medial-lateral extent. By electron microscopy, unmyelinated, small diameter (0.25 μm) ChAT-immuno-reactive axons were observed interspersed among numerous other non-immunoreactive axons in the SNc. ChAT-immunoreactive synaptic endings were observed in juxtaposition to small caliber (0.5 μm) non-immunoreactive dendrites, and contained numerous spheroidal synaptic vesicles and occasional mitochondria. Synaptic contact zones were characterized by an accumulation of synaptic vesicles along the presynaptic membrane, and a prominent postsynaptic densification producing an asymmetrical pre-/postsynaptic membrane profile typical of excitatory synapses. These findings provide direct evidence for a cholinergic innervation of the SN, and suggest that this input may have an excitatory effect on neuronal elements in the SNc.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00248513
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  • 29
    Electronic Resource
    Electronic Resource
    Changes in microglial cell numbers in the spinal cord dorsal horn following brachial plexus transection in the adult rat (1988)
    Springer
    Experimental brain research 73 (1988), S. 61-68 
    Details
    ISSN: 1432-1106
    Keywords: Microglia ; Dorsal horn ; Nerve lesion ; Light microscopy ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The effect of peripheral nerve transection on the size of the microglial cell population in cytoarchitecturally distinct regions of the spinal cord dorsal horn of rats was evaluated at selected intervals 2 through 35 days after unilateral brachial plexotomy. The identification of cells was verified by electron microscopic examination of a representative random sample of cells included in the counts. Microglial cell numbers were increased in laminae I, II as well as the arbitrarily defined deeper laminae 3.5 days after surgery. Although microglial cell numbers in laminae I were within normal range 35 days after axotomy, those of the more ventrally located laminae remained significantly greater than control values for the duration of the experimental period. These findings demonstrate that: 1) microglial cell proliferation in the dorsal horn is an early event in the central changes that are attendant to peripheral nerve injury 2) the time course of the response varies in cytoarchitecturally different regions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00279661
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  • 30
    Electronic Resource
    Electronic Resource
    Hypertension and nephrotoxic lesions in rats 1 year after prenatal exposure to gentamicin (1988)
    Springer
    Archives of toxicology 62 (1988), S. 274-284 
    Details
    ISSN: 1432-0738
    Keywords: Gentamicin ; Postnatal development ; Blood pressure ; Embryotoxicity ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Using the aminoglycoside antibiotic gentamicin persistant functional and morphological changes were induced prenatally in the rat kidney. After 6 days of s.c. treatment (110 mg gentamicin/kg body wt) from day 10 to 15 of pregnancy complete resorption was noticed in 8 of the 14 treated animals. Fifty-three newborn were obtained from six dams. One year later only 26 rats (16 male, 10 female) were still alive. The systolic arterial pressure of the female offspring was significantly increased (139±15 mm Hg versus 112±9 mmHg) compared with controls. No statistically significant effect could be noticed in the male offspring (128±5 mm Hg versus 118±21 mm Hg). Corresponding results were obtained from analysis of urea plasma concentrations. Another cohort of pregnant rats received daily injections of gentamicin from day 15 to 20 of pregnancy (110 mg/kg body wt s.c.). In this group 59 newborn from a total of 109 died within the first 5 days after birth. Six litters were observed postnatally. One year after birth the following blood pressure values were determined: 122±4 mm Hg (male) and 132±17 mm Hg (female). Urea plasma concentrations were significantly higher in female, but not in male, offspring. Light and electron microscopic inspection revealed pathological changes in the kidneys of the female offspring only. The degree of maternal kidney damage — which shows considerable variations — was monitored during the treatment period. For this purpose the plasma gentamicin and urea concentrations were measured on 3 days of treatment in all of the pregnant animals. The postnatal data (mortality, blood pressure, and urea plasma concentrations) show a correlation to the degree of maternal kidney impairment.
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    URL: http://dx.doi.org/10.1007/BF00332487
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  • 31
    Electronic Resource
    Electronic Resource
    Adult-onset rod disease with abundant intranuclear rods (1988)
    Springer
    Journal of neurology 235 (1988), S. 343-347 
    Details
    ISSN: 1432-1459
    Keywords: Adult-onset rod disease ; Nemaline myopathy ; α-Actinin ; Immunohistochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The third case of adult-onset rod disease (nemaline myopathy) with abundant myofibrillar as well as intranuclear rods is described. The 61-year-old woman suffered from progressive weakness of proximal extremities and of the neck, mimicking polymyositis. Muscle biopsy revealed a striking myopathic pattern, with intranuclear rods occurring in 31% of the fibres. On light and electron microscopy and by immunohistochemical study, the rods differed from myofibrillar rods. The absence of α-actinin in intranuclear rods suggests an enhanced readiness of actin filaments to bind to diverse proteins, instead of overproduction of α-actinin as the pathogenetic basis of the rod formation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00314229
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  • 32
    Electronic Resource
    Electronic Resource
    Immunocytochemical localization of two key enzymes of the 2-hydroxyglutarate pathway of glutamate fermentation in Acidaminococcus fermentans (1988)
    Springer
    Archives of microbiology 150 (1988), S. 504-508 
    Details
    ISSN: 1432-072X
    Keywords: Acidaminococcus fermentans ; Glutamate fermentation ; Electron microscopy ; Immunocytochemistry ; Post-embedding labelling ; Antibody-gold complexes ; Protein A-gold complexes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have investigated the in situ location of glutaconyl-CoA decarboxylase and 2-htdroxyglutaryl-CoA dehydratase in Acidaminococcus fermentans using the antibody-gold and protein A-gold techniques carried out as a post-embedding immunoelectron microscopic procedure. Polyclonal antisera were raised in rabbits against homogeneous fractions of the enzymes. Anaerobically grown cells of A. fermentans of the late exponential growth phase were fixed with 0.2% glutaraldehyde and 0.3% formaldehyde (final concentrations) in the growth medium. Dehydration of the cells was achieved with methanol. The cells were embedded in the low temperature embedding resin Lowicryl K4M. The markers indicative for antigenic sites of the two enzymes unequivocally demonstrate that the sodium pump glutaconyl-CoA decarboxylase is located at the cell periphery being a membrane-bound enzyme as expected whereas 2-hydroxyglutaryl-CoA dehydratase is a soluble cytoplasmic enzyme.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00422295
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    Electronic Resource
    Electronic Resource
    The production and utilization of intermediary elemental sulfur during the oxidation of reduced sulfur compounds by Thiobacillus ferrooxidans (1988)
    Springer
    Archives of microbiology 150 (1988), S. 574-579 
    Details
    ISSN: 1432-072X
    Keywords: Thiobacillus ferrooxidans ; Sulfur production ; Sulfur oxidation ; Inhibitors ; Uncouplers ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The intermediary production of elemental sulfur during the microbial oxidation of reduced sulfur compounds has frequently been reported. Thiobacillus ferrooxidans, an acidophilic chemolithoautotroph, was found to produce an insoluble sulfur compound, primarily elemental sulfur, during the oxidation of thiosulfate, trithionate, tetrathionate and sulfide. This was confirmed by light and electron microscopy. Sulfur was produced from sulfide by an oxidative step, while the production from tetrathionate was initiated by a hydrolytic step, probably followed by a series of chemical reactions. The oxidation of intermediary sulfur was severely inhibited by sulfhydryl-binding reagents such as N-ethylmaleimide, by the addition of uncouplers or after freezing and thawing of the cells, which probably damaged the cell membrane. The mechanisms behind these inhibitions have not yet been clarified. Finally, it was observed that elemental sulfur oxidation by whole cells depended on the medium composition. The absence of sulfate or selenate reduced the sulfur oxidation rate.
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    URL: http://dx.doi.org/10.1007/BF00408252
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    Electronic Resource
    Electronic Resource
    Structure of the mitochondrial genome of Beta vulgaris L. (1988)
    Springer
    Theoretical and applied genetics 76 (1988), S. 753-759 
    Details
    ISSN: 1432-2242
    Keywords: Plant mitochondrial genome ; Minicircle DNA ; Electron microscopy ; Beta vulgaris L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The structure of mitochondrial DNA (mt-DNA) from sugarbeet (Beta vulgaris L.) has been studied by biochemical methods and electron microscopy. It was found to be complex multipartite consisting of two main classes of molecules: high molecules weight (HMW) mtDNA and low molecular weight (LMW) mtDNA. The HMW mtDNA consists of rosette-like structures and globules resembling chromomeres (150–200nm). A typical rosette has a protein core and radially stemming closed DNA loops (from 0.6-1.5 μm). The number of loops in a rosette varies from 16–30. The bulk of HMW mtDNAs are represented by interconnected rosettes (total contour length about 130–160 μm, 403–496 kbp). Such large circular DNAs may be evidence of the master chromosome arrangement of the sugarbeet genome. Globules and rosettes are interconnected by thick and thin DNA fibrils, along which nucleosome- and nucleomere-like structures are distributed. The LWM mtDNA is composed of two groups of supercoiled circular molecules, 0,2–1.5 μm and 0.02–0.05 μm in size. Electrophoretic analysis demonstrated that LWM mtDNA is represented by minicircle plasmid-like DNA molecules of 1.3, 1.4 and 1.6 kbp.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00303522
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  • 35
    Electronic Resource
    Electronic Resource
    Distribution of mitochondrial plasmid-like DNA in cultivated rice (Oryza sativa L.) and its relationship with varietal groups (1988)
    Springer
    Theoretical and applied genetics 76 (1988), S. 809-814 
    Details
    ISSN: 1432-2242
    Keywords: Electron microscopy ; Genetic variation ; Mitochondrial DNA ; Oryza sativa L. ; Plasmid-like DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mitochondrial (mt) plasmid-like DNA was found in most of more than 100 rice cultivars (Oryza sativa L.) by the use of 0.7% agarose gel electrophoresis (AGE). The DNA varied in molecular weight and number. By electron microscopy, small circular DNAs of different sizes could be detected in addition to the DNAs of high molecular weight, even in cultivars in which mt plasmid-like DNA was not detected by AGE. The detection of the mt plasmid-like DNAs by AGE did not depend on their presence or absence, but on their high stoichiometry. The relationship between cytoplasms with mt plasmid-like DNAs and varietal (for example, Indica rice) groups was close. The geographical distribution of cytoplasms is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00273665
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  • 36
    Electronic Resource
    Electronic Resource
    Localization of 28 kDa calbindin in human odontoblasts (1988)
    Springer
    Cell & tissue research 254 (1988), S. 341-346 
    Details
    ISSN: 1432-0878
    Keywords: Odontoblast ; Calbindin ; Immunohistochemistry ; Electron microscopy ; Teeth ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence of 28 kDa calbindin in human odontoblasts was studied by use of specific antibodies raised against chick duodenal 28 kDa calbindin, in immunofluorescence, immuno-peroxidase, and electron-microscopic labelling experiments. The calbindin-like protein was detected mainly in the cytoplasm of odontoblast cell bodies, in their processes and occasionally in their nuclei. Correspondingly, at the ultrastructural level, immunoreactive material was associated with the cytosol, microfilaments and cilia. These findings suggest that human odontoblasts express a 28 kDa vitamin D-dependent calcium-binding protein, unlike those of rats and mice in which ameloblasts are the only cells immunoreactive for the protein.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00225806
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  • 37
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    Electronic Resource
    Development of follicular dendritic cells in lymph nodes of B-cell-depleted mice (1988)
    Springer
    Cell & tissue research 254 (1988), S. 449-454 
    Details
    ISSN: 1432-0878
    Keywords: Follicular dendritic cells ; Lymph nodes ; Electron microscopy ; Nude mice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We have studied follicular dendritic cells (FDC) in lymph nodes of normal and thymus dysgeneic nude mice depleted of B-cells by chronic treatment with anti-IgM antibodies. We found that B cell depletion was accompanied by the absence of mature FDC as defined morphologically at the ultrastructural level. Only precursor FDC (p-FDC) could be demonstrated. Upon release of B-cell suppression, the repopulation of lymph nodes with B-cells was associated with the reappearance of fully differentiated FDC in primary follicles of nude mice and in secondary follicles of T-cell competent mice. We conclude that mature B-cells and/or B-cell products are required for the development of mature follicular dendritic cells in the mouse lymph node.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00225818
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    Electronic Resource
    Orthogonal arrays of particles in non-pigmented cells of rat ciliary epithelium: Relation to distribution of filipin- and digitonin-induced alterations of the basolateral membrane (1988)
    Springer
    Cell & tissue research 252 (1988), S. 165-173 
    Details
    ISSN: 1432-0878
    Keywords: Ciliary epithelium ; Orthogonal arrays of particles ; Filipin ; Freeze-fracture ; Electron microscopy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary It has been suggested that orthogonal arrays of particles may increase the rigidity of plasma membrane, as does cholesterol. Therefore, using freeze-fractured non- pigmented ciliary epithelium, the distribution of such arrays was compared to the distribution of membrane deformations induced by the sterol-probes filipin and digitonin in different domains of the basolateral plasma membrane. The distribution of orthogonal arrays of particles was homogeneous between different regions of the basolateral membrane of the non-pigmented ciliary epithelium, while the number of filipin-induced alterations was nearly 4 times higher in the membrane domains not in contact with the basal lamina than in domains in contact with it. Contrary to the homogeneous distribution of arrays, digitonin-induced deformations also differed markedly in these two basolateral membrane domains. Considering that a marked positive response to sterol probes implies a high sterol content, we conclude that orthogonal arrays of particles can occur in plasma membrane regions well-provided with cholesterol and not in direct contact with the basal lamina. Other possible roles of these arrays are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00213838
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  • 39
    Electronic Resource
    Electronic Resource
    Ultrastructural localization of immunolabeled substance P and methionine-enkephalin-octapeptide in the surface layer of the dorsal horn of rat spinal cord (1988)
    Springer
    Cell & tissue research 253 (1988), S. 55-60 
    Details
    ISSN: 1432-0878
    Keywords: Substance P ; Enkephalin ; Spinal dorsal horn ; Immunocytochemistry ; Electron microscopy ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A preembedding dual immunolabeling technique and electron microscopy were utilized to demonstrate the localization of immunoreactive substance P and methionine-enkephalin-octapeptide (Enk-8) in ultrathin sections of the surface layer (laminae I and II) of rat spinal dorsal horn. The immunoreaction of Enk-8 was visualized as goldtoned silver particles and that of substance P as diaminobenzidine reaction products. Axonal terminals with immunoreactive substance P, and also unlabeled axonal terminals, formed synaptic junctions with the perikarya and dendritic processes of Enk-8-containing neurons. Dendritic profiles immunolabeled for substance P were synaptically linked with unlabeled axons but not with Enk-8-positive ones. Furthermore, it was found that Enk-8 axons and substance P axons terminated synaptically in juxtaposition to one another on the same immunonegative dendrites. Among the Enk-8-containing neurons axonal profiles also appeared to be synaptically associated with immunoreactive Enk-8 dendritic processes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00221739
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    Electronic Resource
    Electronic Resource
    Light- and electron-microscopic immunocytochemistry of peptidergic neurons innervating thoracico-abdominal neurohaemal areas in the blowfly (1988)
    Springer
    Cell & tissue research 253 (1988), S. 583-595 
    Details
    ISSN: 1432-0878
    Keywords: Bioactive peptides ; Coexistence of peptides ; Immunocytochemistry ; Electron microscopy ; Insect nervous system ; Calliphora erythrocephala, C. vomitoria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Ventral thoracic neurosecretory cells (VTNCs) of the blowflies, Calliphora erythrocephala and C. vomitoria, innervating thoracic neuropil and the dorsal neural sheath of the thoracico-abdominal ganglion have been shown to be immunoreactive to a variety of mammalian peptide antisera. In the neural sheath the VTNC terminals form an extensive neurohaemal network that is especially dense over the abdominal ganglia. The same areas are invaded by separate, ut overlapping serotonin-immunoreactive (5-HT-IR) projections derived from neuronal cell bodies in the suboesophageal ganglion. Immunocytochemical studies with different antisera, applied to adjacent sections at the lightmicroscopic level, combined with extensive cross-absorption tests, suggest that the perikarya of the VTNCs contain co-localized peptides related to gastrin/cholecystokinin (CCK), bovine pancreatic polypeptide (PP), Met- and Leuenkephalin and Met-enk-Arg6-Phe7 (Met-enk-RF). Electron-microscopic immunogold-labeling shows that some of the terminals in the dorsal sheath react with several of the individual peptide antisera, whilst others with similar cytology are non-immunoreactive. In the same region, separate terminals with different cytological characteristics contain 5-HT-IR. Both 5-HT-IR and peptidergic terminals are localized outside the cellular perineurium beneath the acellular permeable sheath adjacent to the haemocoel. Hence, we propose that various bioactive substances may be released from thoracic neurosecretory neurons into the circulating haemolymph to act on peripheral targets. The same neurons may also interact by synaptic or modulatory action in the CNS in different neuropil regions of the thoracic ganglion.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00219749
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    Electronic Resource
    Electronic Resource
    The efferent innervation of the genital chamber by an identified serotonergic neuron in the female cricket Acheta domestica (1988)
    Springer
    Cell & tissue research 252 (1988), S. 449-457 
    Details
    ISSN: 1432-0878
    Keywords: Serotonin (5-HT) ; Genital chamber ; Immuno-cytochemistry ; Electron microscopy ; Cricket, Acheta domestica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The serotonergic innervation of the genital chamber of the female cricket, Acheta domestica, has been investigated applying anti-serotonin (5-HT) immunocyto-chemistry at both light- and electron-microscopic levels as well as using conventional electron microscopy. Whole mount and pre-embedding chopper techniques of immuno-cytochemistry reveal a dense 5-HT-immunoreactive network of varicose fibers in the musculature of the genital chamber. All of these immunoreactive fibers originate from the efferent serotonergic neuron projecting through the nerve 8v to the genital chamber (Hustert and Topel 1986; Elekes et al. 1987). At the electron-microscopic level, 5-HT-immunoreactive nerve terminals, which contain small (50–60 nm) and large (∼ 100 nm) agranular vesicles as well as granular vesicles (∼100nm), contact the muscle fibers or the sarcoplasmic processes without establishing specialized neuromuscular connections. In addition to the 5-HT-immunoreactive axons, two types of immunonegative axons can also be found in the musculature. By use of conventional electron microscopy, three ultrastructurally distinct types of axon processes can be observed, one of which resembles 5-HT-immunoreactive axons. While the majority of the varicosities do not synapse on the muscle fibers, terminals containing small (50–60 nm) agranular vesicles occasionally form specialized neuromuscular contacts. It is suggested that the 5-HTergic innervation plays a non-synaptic modulatory role in the regulation circular musculature in the genital chamber of the cricket, while the musculature as a whole may be influenced by both synaptic and modulatory mechanisms.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00214388
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  • 42
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    Electronic Resource
    Atrial-specific granules in the hearts of normal and water-deprived rats (1988)
    Springer
    Cell & tissue research 253 (1988), S. 547-552 
    Details
    ISSN: 1432-0878
    Keywords: Atrial-specific granule ; Atrial natriuretic polypeptide ; Water deprivation ; Immunohistochemistry ; Electron microscopy ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The morphology of atrial-specific granules, which contain atrial natriuretic polypeptide (ANP), was studied in the cardiac tissue of untreated controls and water-deprived rats by means of conventional and immunoelectron microscopy. Immature secretory vesicles or granules appeared to become buded off from the Golgi cisternae and then fused to form specific A-granules. An electron-dense plate with a fuzzy coat was frequently found on the limiting membrane at the end of such fusion. Pale specific B-granules, which were less electron-dense, larger, and more granular than A-granules, were found in small numbers in the left atrial cardiocytes, but rarely in the right ones. Very pale granules with a less granular matrix, considered to be B-type granules which had lost their electron-density, and which had less immunoreactivity for ANP, were numerous in the cardiac tissue after water deprivation. This morphological change, which is interpreted as an indication of granule degradation, was in agreement with the noted increase of natriuretic activity in the atrial tissue of water-deprived specimens.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00219745
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    Electronic Resource
    Epidermal growth factor (EGF)-induced morphological changes in the basement membrane of chick embryonic skin (1988)
    Springer
    Cell & tissue research 254 (1988), S. 481-485 
    Details
    ISSN: 1432-0878
    Keywords: Embryonic skin ; Epidermal growth factor ; Basement membrane ; Electron microscopy ; Chick
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effect of epidermal growth factor (EGF) on the basement membrane structure of chick embryonic skin cultured in a chemically defined medium (BGJb) containing 20 mM hydrocortisone, and EGF at 10, 50, or 100 ng/ml supplemented with 5% delipidized fetal calf serum, was examined by electron microscopy. During development of the epidermis in vitro, EGF (100 ng/ml) caused striking changes to occur in the basement membrane structure and in the keratinization process. The basement membrane frequently became discontinuous with many gaps apparent in section, and occasionally became folded following detachment from the basal surface of the epidermis and protruded into the underlying dermis. In the basal and intermediate cells of EGF-treated epidermis, tonofilament bundles were decreased in number, while desmosomes and hemidesmosomes revealed no significant changes in morphology.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00226497
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    Electronic Resource
    Unifying principles in intermediate filament (IF) structure and assembly (1988)
    Springer
    Protoplasma 145 (1988), S. 73-81 
    Details
    ISSN: 1615-6102
    Keywords: Intermediate filament structure ; Intermediate filament assembly ; Desmin ; Keratins ; Neurofilaments ; Nuclear lamins ; Cytoskeleton ; Electron microscopy ; Polymerization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01349341
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  • 45
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    Electronic Resource
    Evaluation of 111In labelled white blood cells by in vitro functional tests and electron microscopy (1988)
    Springer
    European journal of nuclear medicine 14 (1988), S. 159-164 
    Details
    ISSN: 1619-7089
    Keywords: 111In-white blood cells ; Chemotaxis ; Phagocytosis ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We have studied the influence of granulocyte labelling with commercially available 111In-oxine, tropolone (trop) or home made 111In-Mercapto pyridine (Merc) prepared by the method of Thakur (1985) on the cell structure by electron microscopy and on the cell function by enzymatic tests, random migration, chemotaxis, phagocytosis and bactericidal activity. The granulocytes were labelled with 400 μCi 111In-oxine in saline or 111In-trop or Merc in plasma. The effect of the chelating agents with and without addition of the tracer was studied (n=4) with varying concentrations: 5–10 μg/ml oxine, 10–160 μg/ml trop and 1–4 μg/ml Merc. Chemotaxis and random migration were not affected by 111In-trop and clearly supressed by 111In-oxine and Merc; the other tests were normal. The cell structure was disturbed by Merc. The labelling efficiency was excellent with oxine (90%), acceptable with trop (30%–80%) and poor with Merc (10%–25%). Without 111In, chemotaxis and random migration were normal up to a concentration of 80 μg/ml trop, 8.5 μg/ml oxine and 1 μg/ml Merc. With addition of 111In, chemotaxis and random migration were unaffected up to 80 gmg/ml by trop and markedly supressed by Merc and oxine. It is concluded that labelling with 111In-trop assures intact cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00293542
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    Electronic Resource
    The barrier-like role of activated connective tissue against the spread of supraglottic laryngeal carcinoma (1988)
    Springer
    European archives of oto-rhino-laryngology and head & neck 245 (1988), S. 151-154 
    Details
    ISSN: 1434-4726
    Keywords: Supraglottic carcinoma ; Light microscopy ; Electron microscopy ; Activated connective tissue barrier
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Light microscopic investigations of whole organ serial sections of 15 laryngectomy specimens with supraglottic carcinomas and comparative electron microscopic studies of the upper and lower poles of the tumors were carried out. The ultrastructure of different poles of the same supraglottic tumors was investigated and showed a more progressive fibrillogenesis with an activation of the connective tissue elements of the border on the lower pole of the malignancy. A significant role is attributed to the barrierforming feature of activated connective tissue with respect to the inhibition of the extension of tumor into the glottic region.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00464017
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  • 47
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    Electronic Resource
    Ultrastructural localization of Ca2+-binding sites in the spiral limbus, the stria vascularis and Reissner's membrane of the guinea pig (1988)
    Springer
    European archives of oto-rhino-laryngology and head & neck 245 (1988), S. 279-283 
    Details
    ISSN: 1434-4726
    Keywords: Calcium ; Reissner's membrane ; Stria vascularis ; Limbus ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Cations were precipitated with potassium antimonate in the cochlea of the guinea pig and electron microscopy was used to analyze the distribution of the formed reaction products. Differences in precipitate density between neighboring cells in Reissner's membrane, in the stria vascularis and in the limbus are described. Electron spectroscopic imaging was performed to obtain information about the spatial distribution of the precipitates and their elemental composition.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00464630
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  • 48
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    Electronic Resource
    Intermolecular localization of epitopes within an oligomeric protein by immunoelectron microscopy and image processing (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 161-183 
    Details
    ISSN: 0887-3585
    Keywords: hemocyanin ; correspondence analysis ; monoclonal antibodies ; electron microscopy ; images analysis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Three epitopes have been localized by immunoelectron microscopy on subunits Aa6 of the 4 × 6-meric hemocyanin of the scorpion Androctonus australis. Soluble immunocomplexes composed of monoclonal antibodies and of native hemocyanin were purified, negatively stained with uranyle acetate by the single-layer technique, and examined under the electron microscope (EM). The molecule images were digitized, aligned, and submitted to correspondence analysis according to the method of Van Heel and Frank (Ultramicroscopy 6: 187-194, 1981). A high-precision localization of the attachment point of the Fab arm to the antigen was achieved through a careful analysis of the average images. This method easily allowed the discrimination of epitopes located in different domains (Mr 20 kDa) of the same subunit. Nonoverlapping epitopes located in the same structural domain of subunit Aa6 could be distinguished by the stain exclusion patterns of their Fab arms. The method is general and may be used for epitope mapping in any antigen producing definite EM views.
    Additional Material: 12 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340030305
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  • 49
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    Electronic Resource
    Crystallization and preliminary X-ray diffraction study of a protein with a high potential rubredoxin center and a hemerythrin-type Fe center (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 184-186 
    Details
    ISSN: 0887-3585
    Keywords: new Fe-protein ; rubredoxin ; hemerythrin ; crystals ; X-ray diffraction ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A newly discovered iron-containing protein, isolated from the bacterium Desulfovibrio vulgaris (Hildenborough, NCIB 8303), has been crystallized. The molecule appears to be a dimer of mass 44kDa. This protein has iron centers with spectroscopic similarities to those in rubredoxins and in hemerythrins.The X-ray diffraction shows symmetry consistent with space group I222 or I212121. Cell parameters are a = 49.2 Å, b = 81.3 Å, c= 100.1 Å, and α, β, γ = 90°. X-ray diffraction data have been collected to 3.0 Å, and a search for useful heavy atom derivatives is in progress for the analysis of the crystal structure of this Fe-protein.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340030306
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  • 50
    Electronic Resource
    Electronic Resource
    Lecithin: Cholesterol acyltransferase activation by synthetic amphipathic peptides (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 187-198 
    Details
    ISSN: 0887-3585
    Keywords: amphipathic peptide ; liposomes ; peptide ; serum apolipoproteins ; synthetic ; LCAT ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The amphipathic helical theory of Segrest and colleagues (FEBS Lett.: 38: 247-253, 1974) proposes that the lipid-binding segments of serum apolipoproteins are in an alpha helical conformation. Furthermore the helices have a hydrophobic face and a hydrophilic face with a specific distribution of positively and negatively charged residues. The importance of the pattern of the charged residues in the lipid binding and lecithin:cholesterol acyltransferase (LCAT) activation by the segments is still debated. We designed a 30-residue peptide, GALA, which in the alpha helical conformation hs a hydrophilic face composed of glutamic acid residues (Sabbarao et al.: Biochemistry 26: 2964-2972, 1987). GALA behaves like the serum apolipoproteins in its interaction with dimyristoylphospatidylcholine (DMPC) at neutral pH; the amino terminal tryptophan of GALA undergoes a blue shift in its fluorescence emission spectrum, and the circular dichroism (CD) spectrum indicates that GALA acquires alpha helical structure in the presence of DMPC. A DMPC-GALA:19/1 (molar ratio) complex can be isolated by gel-permeation chromatography. This complex has a discoidal structure with the approximate dimensions of 44-Å diameter. GALA edge thickness and a 170- to 350-Å diameter. GALA activates LCAT with DMPC but not with unsaturated phospholipids as the substrate. The apparent partition coefficient of GALA into DMPC vesicles is 100-fold larger than into egg phosphatidlylcholine vesicles. The interaction of GALA with unsaturated lipids at neutral pH is so weak that no detectable change in the spectroscopic properties of GALA or the structure of the liposomes can be detected under the conditions used here. The sequence of GALA differs from previously studied model Apo A1 peptides by the absence of positively charged residues on the hydrophilic face. This indicates that positive charges in Apo A1-like peptides are not required in order to form discoidal structures with saturated phospholipids or to activate LCAT with such lipid substrates.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340030307
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    In memorium: Emil Thomas Kaiser, 1938-1988 (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. i 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340040102
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  • 52
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    Photochemical crosslinking of bacteriophage T4 single-stranded DNA-binding protein (gp32) to oligo-p(dT)8: Identification of phenylalanine-183 as the site of crosslinking (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 1-6 
    Details
    ISSN: 0887-3585
    Keywords: single-standard DNA-binding protein ; protein-nucleic acid interactions ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Using ultraviolet light, both the 33,000-dalton single-standard DNA-binding protein from T4 bacteriophage (gp32)as well as a 25,000-dalton limited trypsin cleavage product of gp32 (core gp32*) that retains high affinity for single-stranded DNA can be crosslinked to an oligodeoxynucleotide, p(dT)8. After photolysis, a single tryptic peptide crosslinked to p(dT)8 was isolated by anion-exchange high-performance liquid chromatography. Gas-phase sequencing of this modified peptide gave the following sequence: Gln-Val-Ser-Gly-(X)-Ser-Asn-Tyr-Asp-Glu-Ser-Lys, which corresponds to residues 179-190 in gp32. Based on the absence of the expected phenylthiohydantoin derivative of phenylalanine 183 at cycle 5 (X) we infer that crosslinking has occurred at this position and that phenylalanine 183 is at the interface of the gp32:P(dT)8 complex in an orientation that allows covalent bond formation with the thymine radical produced by ultraviolet irradiation.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340040103
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  • 53
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    Structure-function relationships in 3-phosphoglycerate kinase: Role of the carboxy-terminal peptide (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 56-62 
    Details
    ISSN: 0887-3585
    Keywords: Protein engineering ; mutagenesis ; enzyme catalysis ; conformational changes ; domain movement ; hinge bending ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Yeast 3-phosphoglycerate kinase (PGK) is a monomeric enzyme (Mr ∼ 45,000) composed of two globular domains. Each domain corresponds approximately to the amino- and carboxyterminal halves of the polypeptide chain. The carboxy-terminal end extends over the interdomain “hinge” region and packs against the amino-terminal domain. It has been proposed that domain movement, resulting in closure of the active site left, is essential for the catalytic of PGK. Large-scale conformational changes have also been postulated to explain activation of the enzyme by sulfate ions. Using site-specific mutagenesis, we have removed a 15-amino-acid carboxy-terminal fragment, in order to probe its role in the substrate- and sulfate-induced conformational changes. The truncated enzyme exhibited approximately 1% of the activity of native PGK and lost the ability to undergo sulfateinduced activation. The Km for ATP was essentially unchanged (Km = 0.23mM), whereas the Km value for 3-phosphoglycerate was increased about eightfold (Km = 3.85 mM and 0.50 mM, respectively). These results suggest that the carboxy-terminal segment is important for the mechanism of substrate- and specific-induced conformational transitions. CD spectra and sedimentation velocity measurements indicate that the carboxy-terminal peptide is essential for structural integrity of PGK. The increased susceptibility of the truncated enzyme to thermal inactivation implies that the carboxy-terminal peptide also contributes to the stability of PGK.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340040108
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  • 54
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    Model of a complex between the tetrahemic cytochrome c3 and the ferredoxin I from Desulfovibrio desulfuricans (Norway strain) (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 63-70 
    Details
    ISSN: 0887-3585
    Keywords: molecular graphics ; protein complex ; electron transfer ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A three-dimensional model of an electron-transfer complex between the tetrahemic cytochrome c3 and the ferredoxin I from the sulfatereducing bacterium Desulfovibrio desulfuricans (Norway strain) has been generated through computer graphics methods. The model is based on the known X-ray structure of the cytochrome and on a model of the ferredoxin that has been derived through computer graphics modeling and energy minimization methods, from the X-ray structure of the homologous ferredoxin from Peptococcus aerogenes. Four possible models of interaction between the two molecules were examined by bringing in close proximity each of the four hemes and the redox center (4Fe-4S) of the ferredoxin and by optimizing the ion pairs interactions. One of these models shows by far the “best” structure in terms of charges, interactions, and complementary f the topology of the contact surfaces. In this complex, the distance between the iron atoms of the ferredoxin redox center and the hemic iron atom is 11.8 Å, which compares well with those found between redox centers in other complexes. The contact surface area between the two molecules is 170 Å2.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340040109
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  • 55
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    Masthead (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988) 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340040201
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  • 56
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    Crystallization and preliminary investigation of single crystals of deoxyuidine triphosphate nucleotidohydrolase from Escherichia coli (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 71-75 
    Details
    ISSN: 0887-3585
    Keywords: dUTPase ; nucleotide binding enzyme ; X-ray crystallography ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Deoxyuridine triphosphate nucleotidohydrolase (dUTPase), an enzyme in the nucleotide metabolism that is a pyrophosphatase hydrolyzing dUTP, has been crystallized. The crystals belong to the trigonal space group R3 and diffract beyond 2 Å. The native dUTPase crystals and a mercury derivative are stable in the X-ray beam and are suitable for a high resolution X-ray structure analysis.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340040110
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    Crystallographic studies of inhibitor binding sites in human carbonic anhydrase II: A pentacoordinated binding of the SCN- ion to the zinc at high pH (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 283-293 
    Details
    ISSN: 0887-3585
    Keywords: crystallography ; structure ; refinement ; sulfonamide ; thiocyanate ; mercury ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The binding of four inhibitors - mercuric ion, 3-acetoxymercuri-4-aminobenzenesulfonamide (AMS), acetazolamide (Diamox), and thiocyanate ion - to human carbonic anhydrase II (HCA II) has been studied with X-ray crystallography.The binding of mercury to HCA II at pH 7.0 has been investigated at 3.1 Å resolution. Mercuric ions are observed at both nitrogens in the His-64 ring. One of these sites is pointing toward the zinc ion. The only other binding site for mercury is at Cys-206.The binding of the two sulfonamide inhibitors AMS and Diamox, has been reinvestigated at 2.0 and 3.0 Å, respectively. Only the nitrogen of the sulfonamide group binds to the zinc ion replacing the hydroxyl ion. The sulfonamide oxygen closet to the zinc ion is 3.1 Å away. Thus the tetrahedral geometry of the zinc is retained, refuting earlier models of a pentacoordinated zinc.The structure of the thiocyanate complex has been investigated at pH 8.5 and the structure has been refined at 1.9 Å resolution using the least-squares refinement program PROLSQ. The crystallographic R factor is 17.6%. The zinc ion is pentacoordinated with the anion as well as a water molecule bound in addition to the three histidine residues. The nitrogen atom of the SCN- ion is 1.9 Å from the zinc ion but shifted 1.3 Å with respect to the hydroxyl ion in the native structure and at van der Waals' distance from the Oγl atom of Thr-199. This is due to the inability of the Oγl atom of Thr-199 to serve as a hydrogen bond donor, thus repelling the nonprotonated nitrogen. The SCN- molecule reaches into the deep end of the active site cavity where the sulfur atom has displaced the so-called “deep” water molecule of the native enzyme. The zinc-bound water molecule is 2.2 Å from the zinc ion and 2.4 Å from the SCN- nitrogen. In addition, this water is hydrogen bonded to the Oγl atom of Thr-199 and to another water molecule.We have observed that solvent and inhibitor molecules have three possible binding sites on the zinc ion and their significance for the catalysis and inhibition of HCA II will be discussed. All available crystallographic data are consistent with a proposed catalytic mechanism in which both the OH moiety and one oxygen of the substrate HCO3- ion are ligated to the zinc ion.
    Additional Material: 9 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340040407
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    Further experimental studies of the disulfide folding transition of ribonuclease A (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 251-261 
    Details
    ISSN: 0887-3585
    Keywords: protein folding kinetics ; disulfide bonds ; thiol-disulfide exchange ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Two very different mechanisms of folding have been proposed from experimental studies of disulfide formation in reduced ribonuclease A. (1) A pathway in which the rate-limiting step separates fully folded protein from all other disulfide intermediates and occurs solely in three-disulfide intermediates. (2) A multiple pathway mechanism with different rate-limiting steps for each pathway. The various rate-limiting steps involve disulfide breakage, formation, and rearrangement in intermediates with one, two, three, and four protein disulfides. To distinguish between these two mechanisms, we have carried out further studies of both unfolding and refolding.Refolding of reduced ribonuclease A requires three-disulfide intermediates to accumulate; negligible refolding occurs when only the nearly random one- and two-disulfide intermediate species are populated. Therefore, no rate-limiting steps of the type postulated in mechanism (2) occur in intermediates with one and two protein disulfides. Unfolding and disulfide reduction is an all-or-none process; no disulfide intermediates accumulate to detectable to detectable levels or precede the rate-limiting step. Mechanism (2) requires that such intermediates precede the rate-limiting step and accumulate to substantial levels.The different proposal were shown not to result from the use of different solution conditions or disulfide reagents; the two sets of data are not inconsistent. Instead, the inappropriate mechanism (2) resulted from an incorrect kinetic analysis and misinterpretation of the kinetics of disulfide formation are breakage.
    Additional Material: 6 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340040404
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    Masthead (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988) 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340030201
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    Conformational characteristics of the complete cequence of group A streptococcal M6 protein (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 60-69 
    Details
    ISSN: 0887-3585
    Keywords: coiled-coli ; alpha-helix ; antiphagocytic ; heptad ; antigenic variation ; sequence repeats ; cell wall protein ; intermediate filaments ; myosin ; tropomyosin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: M protein is considered a virulence determinant on the streptococcal cell wall by virtue of its ability to allow the organism to resist attack by human neutrophils. The complete DNA sequence of the M6 gene from streptococcal strain D471 has allowed, for the first time, the study of the structural characteristics of the amino acid sequence of an entire M protein molecule. Predictive secondary structural analysis revealed that the majority of this fibrillar molecule exhibits strong alpha-helical potential and that, except for the ends, nonpolar residues in the central region of the molecule exhibit the 7-residue periodicity typical for coiled-coil proteins. Differences in this heptad pattern of nonpolar residues allow this central rod region to be divided into three subdomains which correlate essentially with the repeat regions A, B, and C/D in the M6 protein sequence. Alignment of the N-terminal half of the M6 sequence with PepM5, the N-terminal half of the M5 protein, revealed that 42% of the amino acids were identical. The majority of the identities were “core” nonpolar residues of the heptad periodicity which are necessary for the maintenance of the coiled coil. Thus, conservation of structure in a sequence-variable region of these molecules may be biologically significant. Results suggest that serologically different M proteins may be built according to a basic scheme: an extended central coiled-coil rod domain (which may vary in size among strains) flanked by functional end domains.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340030106
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    Identification of structural motifs from protein coordinate data: Secondary structure and first-level supersecondary structure*A FORTRAN version of the program DEFINE_STRUCTURE, which defines the secondary structure and characterizes the first level of supersecondary structure, is available from the authors (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 71-84 
    Details
    ISSN: 0887-3585
    Keywords: Cα coordinates ; distance matrix ; difference distance matrix ; helix axes, strand axes ; interaxial angles ; turns ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A computer program is described that produces a description of the secondary structure and supersecondary structure of a polypeptide chain using the list of alpha carbon coordinates as input. Restricting the term “secondary structure” to the conformation of contiguous segments of the chain, the program determines the initial and final residues in helices, extended strands, sharp turns, and omega loops. This is accomplished through the use of difference distance matrices. The distances in idealized models of the segments are compared with the actual structure, and the differences are evaluated for agreement within preset limits. The program assigns 90-95% of the residues in most proteins to at least one type of secondary elementIn a second step the now-defined helices and strands are idealized as straight line segments, and the axial directions and locations are compiled from the input Cα coordinate list. These data are used to check for moderate curvature in strands and helices, and the secondary structure list is corrected where necessary. The geometric relations between these line segments are then calculated and output as the first level of supersecondary structure. A maximum of six parameters are required for a complete description of the relations between each pair. Frequently a less complete description will suffice, for example just the interaxial separation and angle. Both the secondary structure and one aspect of the supersecondary structure can be displayed in a character matrix analogous to the distance matrix format. This allows a quite accurate two-dimensional display of the three-dimensional structure, and several examples are presentedA procedure for searching for arbitrary substructures in proteins using distance matrices is also described. A search for the DNA binding helix-turnhelix motif in the Protein Data Bank serves as an exampleA further abstraction of the above data can be made in the form of a metamatrix where each diagonal element represents an entire secondary segment rather than a single atom, and the off-diagonal elements contain all the parameters describing their interrelations. Such matrices can be used in a straightforward search for higher levels of supersecondary structure or used in toto as a representation of the entire tertiary structure of the polypeptide chain.
    Additional Material: 5 Ill.
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    Pattern descriptors and the unidentified reading frame 6 human mtDNA dinucleotide-binding site (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 97-101 
    Details
    ISSN: 0887-3585
    Keywords: URF ; nucleotide-binding sites ; pattern descriptor ; computer search ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: In an effort to identify the structural elements essential to a given protein function a new pattern-directed inference system has been developed. It has been employed to identify a potential dinucleotide-binding domain within the human mitochondrial unidentified reading frame 6 product, thereby supporting an earlier study that this gene may encode a NADH dehydrogenase subunit.
    Additional Material: 2 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340030204
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    Lysine/fibrin binding sites of kringles modeled after the structure of kringle 1 of prothrombin (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 85-96 
    Details
    ISSN: 0887-3585
    Keywords: molecular modeling ; energy minimization ; lysine/fibrin binding ; kringle structures ; plasminogen ; tissue plasminogen activator ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The Lys binding site of kringle 1 and 4 (K1 and K4) of plasminogen (PG) has been modeled on the basis of the three-dimensional structure of kringle 1 of prothrombin and 300- and 600-MHz proton nuclear magnetic resonance observations. These structures were then compared to the corresponding regions of modeled kringle 1 and 2 of tissue plasminogen activator (PA). The coordinates of the modeled structures have been refined by energy minimization in the presence and absence of ∊-aminocaproic acid ligand in order basically to remove unacceptable van der Waals contacts. The binding site is characterized by an apparent dipolar surface, the polar parts of which are separated by a hydrophobic region of highly conserved aromatic residues. Zwitterionic ligands such as Lys and ∊-aminocaproic acid form ion pair interactions with Asp55 and Asp57 located on the dipolar surface; the latter are also conserved in all the Lys binding kringles. The cationic center of the dipolar surface is Arg71, in the case of PGK4, and is composed of Arg34 and Arg71 in PGK1. The doubly charged anionic/cationic interaction centers of the latter might account for the larger binding constants of PGK1 for like-ligands but the modeling suggests that PGK4 might be kinetically faster in binding bulkier ligands. The binding site region of PAK2, which also binds Lys, resembles those of PGK1 and PGK4. Since PAK2 lacks both cationic center Arg residues, ligand carboxylate binding appears to be accomplished though an imidazolium ion of His64, which is located just below the outer surface of the kringle.
    Additional Material: 6 Ill.
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  • 64
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    Site-directed mutations altering methyl-accepting residues of a sensory transducer protein (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 102-112 
    Details
    ISSN: 0887-3585
    Keywords: bacterial chemotaxis ; sensory adaptation ; protein modification ; membrane protein ; receptor protein ; transmembrane signalling ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The Trg protein is one of a family of transducer proteins that mediate chemotactic response in Escherichia coli. Transducers are methylaccepting proteins that gain or lose methyl esters on specific glutamyl residues during sensory adaptation. In this study, the significance of multiple sites of methylation on transducer proteins was addressed by using oligonucleotide-directed, site-specific mutagenesis to substitute an alanyl residue at each of the five methyl-accepting sites in Trg. The resulting collection of five mutations, each inactivating a single site, was analyzed for effects on covalent modification at the remaining sites on Trg and for the ability of the altered proteins to mediate sensory adaptation. Most of the alanyl substitutions had substantial biochemical effects, enhancing or reducing methyl-accepting activity of other sites, including one case of activation of a site not methylated in wild-type protein. Analysis of the altered proteins provided explanations for many features of the complex pattern of electrophoretic forms exhibited by Trg. The mutant proteins were less efficient than normal Trg in mediating adaptation. Correlation of biochemical and behavioral data indicated that reduction in the number of methyl-accepting sites on the transducer lengthened the time required to reach an adapted state.
    Additional Material: 10 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340030205
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    Human myeloperoxidase and thyroid peroxidase, two enzymes with separate and distinct physiological functions, are evolutionarily related members of the same gene family (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 113-120 
    Details
    ISSN: 0887-3585
    Keywords: evolution ; proximal histidine ; distal histidine ; heme enzyme ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Human myeloperoxidase and human thyroid peroxidase nucleotide and amino acid sequences were compared. The global similarities of the nucleotide and amino acid sequences are 46% and 44%, respectively. These similarities are most evident within the coding sequence, especially that encoding the myeloperoxidase functional subunits. These results clearly indicate that myeloperoxidase and thyroid peroxidase are members of the same gene family and diverged from a common ancestral gene. The residues at 416 in myeloperoxidase and 407 in thyroid peroxidase were estimated as possible candidates for the proximal histidine residues that link to the iron centers of the enzymes. The primary structures around these histidine residues were compared with those of other known peroxidases. The similarity in this region between the two animal peroxidases (amino acid 396-418 in thyroid peroxidase and 405-427 in myeloperoxidase) is 74%; however, those between the animal peroxidases and other yeast and plant peroxidases are not significantly high, although several conserved features have been observed. The possible location of the distal histidine residues in myeloperoxidase and thyroid peroxidase amino acid sequences are also discussed.
    Additional Material: 4 Ill.
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    Peptide synthesis catalyzed by polyethylene glycol-modified chymotrypsin in organic solvents (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 130-137 
    Details
    ISSN: 0887-3585
    Keywords: peptide synthesis ; chymotrypsin specificity ; polyethylene glycol ; nonaqueous solvents ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Chymotrypsin modified with polyethylene glycol was successfully used for peptide synthesis in organic solvents. The benzene-soluble modified enzyme readily catalyzed both aminolysis of N-benzoyl-L-tyrosine p-nitroanilide and synthesis of N-benzoyl-L-tyrosine butylamide in the presence of trace amounts of water. A quantitative reaction was obtained when either hydrophobic or bulky amides of L- as well as D-amino acids were used as acceptor nucleophiles, while almost no reaction occurred with free amino acids or ester derivativesThe acceptor nucleophile specificity of modified chymotrypsin as a catalyst in the formation of both amide and peptide bonds in organic solvents was quite comparable to that in aqueous solution as well as to that of the leaving group in hydrolysis reactions. By contrast, the substrate specificity of modified chymotrypsin in organic solvents was different from that in water since arginine and lysine esters were found to be as effective as aromatic amino acids to form the acyl-enzyme with subsequent synthesis of a peptide bond.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340030208
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    Vibrational spectroscopy of bacteriorhodopsin mutants: I. Tyrosine-185 protonates and deprotonantes during the photocycle (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 219-229 
    Details
    ISSN: 0887-3585
    Keywords: proton transport ; energy transduction ; purple membrane ; proton wire ; Schiff base counter-ion ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The techniques of FTIR difference spectroscopy and site-directed mutagenesis have been combined to investigate the role of individual tyrosine side chains in the proton-pumping mechanism of bacteriorhodopsin (bR). For each of the 11 possible bR mutants containing a single Tyr→Phe substitution, difference spectra have been obtained for the bR→K and bR→M photoreactions. Only the Tyr-185→Phe mutation results in the disappearance of a set of bands that were previously shown to be due to protonation of a tryosinate during the br→K photoreaction [Rothschild et al.: Proceedings of the National Academy of Sciences of the United states of America 83:347, (1986)]. The Tyr-185→Phe mutation also eliminates a set of bands in the bR→M difference spectrum associated with deprotonation of a Tyr; most of these bands (e.g., positive 1272-cm-1 peak) are completely unaffected by the other ten Tyr→Phe mutations. Thus, tyrosinate-185 gains a proton during the bR→K reaction and loses it again when M is formed. Our FTIR spectra also provide evidence that Tyr-185 interacts with the protonated Schiff base linkage of the retinal chromophore, since the negative C=NH+ stretch band shifts from 1640 cm-1 in the wild type to 1636 cm-1 in the Tyr-185→Phe mutant. A model that is consistent with these results is that Tyr-185 is normally ionized and serves as a counter-ion to the protonated Schiff base. The primary photoisomerization of the chromophore translocates the Schiff base away from Tyr-185, which raises the pKa of the latter group and results in its protonation.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340030403
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    Structural comparison of the prokaryotic ribosomal proteins L7/L12 and L30 (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 243-251 
    Details
    ISSN: 0887-3585
    Keywords: protein evolution ; structural homology ; ribosome structure ; x-ray crystallography ; common motif ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The structure of two prokaryotic ribosomal proteins, the carboxyterminal half of L7/L12 from Escherichia coli (L12CTF) and 1.30 from Bacilus Stearothermophilus display a remarkably similar fold in which alpaha-helices pack onto one side of an antiparallel, three-stranded, beta-pleated sheet. A detailed comparison of the structures by least-squares methods reveals that more than two-thirds of the alpha carbons can be superimposed with a root mean square distance of 2.33 Å. The principal difference is an extra alpha-helix in L12 CTF. The sequences of the proteins display a distinct conservation in regions which are crucial to the common fold, in particular the hydrophobic core. It is proposed that the similarity is a result of divergent evolution.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340030405
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    Crystallographic structure analysis of lamprey hemoglobin from anomalous dispersion of synchrotron radiation (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 77-88 
    Details
    ISSN: 0887-3585
    Keywords: protein structure ; diffraction ; anomalous scattering ; x-ray crystallography ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The molecular structure of lamprey hemoglobin was previously determined and refined by conventional crystallographic analysis. In this study, the structural analysis has been repeated in the course of developing the method of multiwavelength anomalous diffraction (MAD) for phase determination. New experimental and analytical procedures that were devised to perform this determination should have general applicability. These include an experimental design to optimize signal strength and reduce systematic errors, experimental evaluation of anomalous scattering factors, and a least-squares procedure for analyzing the MAD data. MAD phases for the structure at 3Å resolution are as accurate overall as the multiple isomorphous replacement (MIR) phases determined previously.
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    Conformation of alamethicin in phospholipid vesicles: Implications for insertion models (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 89-98 
    Details
    ISSN: 0887-3585
    Keywords: membrane proteins ; channels ; circular dichroism spectroscopy ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The secondary structure of alamethicin, a membrane channel-forming polypeptide, has been examined by circular dichroism spectroscopy to determine the relationship of its conformation in organic solution to its conformation in a membrane-bound state. The spectrum of alamethicin in small unilamellar dimyristoyl phosphatidylcholine vesicles is significantly different from its spectrum in 10% methanol/acetonitrile, the solvent from which it was crystallized (Fox and Richards: Nature 300:325-330, 1982), as well as its spectrum in methanol, the solvent in which NMR studies have been done (Banerjee and Chan: Biochemistry 22:3709-3713, 1983). This suggests that structural models based on studies of the molecule in organic solvents may not be entirely appropriate for the membrane-bound state. To distinguish between different models for channel formation and insertion, two different methods were used to associate the alamethicin with vesicles; in addition, the effect of oligomerization on the conformation of the membrane-bound state was investigated. These studies are consistent with a modified insertion model in which alamethicin monomers, dimers, or trimers associate with the bilayer and then spontaneously oligomerize to form a prechannel with a higher helix content. This aggregate could then “open” upon application of an appropriate gating transmembrane potential.
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    Internal motion of a tryptophan residue in Streptomyces subtilisin inhibitor: Deuterium nuclear magnetic resonance in solution (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 131-136 
    Details
    ISSN: 0887-3585
    Keywords: 2H NMR ; selective deuteration ; tryptophan internal motion ; SSI-subtilisin complex ; protein conformational equilibrium ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Deuterium NMR spectroscopy was used to study internal motions of a deuterium-labeled single tryptophan (Trp) residue (per subunit) of Streptomyces subtilisin inhibitor (SSI) in solution. The free inhibitor with the five ring protons of the Trp replaced with deuterons showed a narrow resonance component (56 Hz) of about one-quarter of the total intensity, in addition to the broad resonance component (about 600 Hz) at 25°C, showing that it exits in an equilibrium mixture of two conformers, in one of which the typtophan side chain is highly mobile. In analogy to the two structures of SSI found in the crystal, these two conformers were attributed to the one in which the contact between the α-lobe and the beta;-lobe of the subunit is tight and the other in which the same contact is loose. When SSI forms a complex with subtilisin BPN′, the broad component becomes invisibly broad, but the narrow component increases with even further narrowing, suggesting that the binding to the enzyme favors the “loose” conformer over the “tight” conformer.
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    Surface interactions of γ-crystallins in the crystal medium in relation to their association in the eye lens (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 137-147 
    Details
    ISSN: 0887-3585
    Keywords: eye lens proteins ; protein association ; crystal packing ; surface area ; homologous proteins ; point mutations ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A comparative study of intermolecular interactions in crystals of two homologous low molecular weight proteins, γ-II and γ-IIIb crystallins, from calf eye lens was carried out. Crystal packings for these proteins are very different: intermolecular contact areas compose about 33% of the total accessible surface area of γ-II as compared with 13% in γ-III. Two key residues seem to be mainly responsible for the differences in protein association in the crystal medium. These are Ser 103 and Leu 155 in γ-II, which are replaced by Met 103 and His 155 in γ-IIIb. A similar substitution of these residues is observed in different gene products of γ-crystallins from a number of vertebrates. This is consistent with the existence of a genetically controlled mechanism for determining intermolecular association of γ-crystallins in the native medium of the lens.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340040207
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    Polar hydrogen positions in proteins: Empirical energy placement and neutron diffraction comparison (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 148-156 
    Details
    ISSN: 0887-3585
    Keywords: Protein structure ; empirical energy ; energy minimization ; molecular dynamics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A method for the prediction of hydrogen positions in proteins is presented. The method is based on the knowledge of the heavy atom positions obtained, for instance, from X-ray crystallography. It employs an energy minimization limited to the environment of the hydrogen atoms bound to a common heavy atom or to a single water molecule. The method is not restricted to proteins and can be applied without modification to nonpolar hydrogens and to nucleic acids. The method has been applied to the neutron diffraction structures of trypsin ribonuclease A, and bovine pancreatic trypsin inhibitor. A comparison of the constructed and the observed hydrogen positions shows few deviations except in situations in which several energetically similar conformations are possible. Analysis of the potential energy of rotation of Lys amino and Ser, Thr, Tyr hydroxyl groups reveals that the conformations of lowest intrinsic torsion energies are statistically favored in both the crystal and the constructed structures.
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    Masthead (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988) 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340040301
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    X-ray and model-building studies on the specificity of the active site of proteinase K (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 157-164 
    Details
    ISSN: 0887-3585
    Keywords: synthetic inhibitors ; serine proteinase crystallography ; active site geometry ; computer graphics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Proteinase K, the extracellular serine endopeptidase (E.C. 3.4.21.14) from the fungus Tritirachium album limber, is homologous to the bacterial subtilisin proteases. The binding geometry of the synthetic inhibitor carbobenzoxy-Ala-Phechloromethyl Ketone to the active site of proteinase K was the first determined from a Fourier synthesis based on synchrotron X-ray diffraction data between 1.8 Å and 5.0 Å resolution. The protein inhibitor complexes was refined by restrained least-squares minimization with the data between 10.0 and 1.8 Å. The final R factor was 19.1% and the model contained 2,018 protein atoms, 28 inhibitors atoms, 125 water molecules, and two Ca2+ ions. The peptides portion of the inhibitor is bound to the active center of proteinase K by means of a three-stranded antiparallel pleated sheet, with the side chain of the phenylalanine located in the P1 site. Model building studies, with lysine replacing phenylalanine in the inhibitor, explain the relatively unspecific catalytic activity of the enzyme.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340040302
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    Structural analysis of the 2.8 Å model of xylose isomerase from Actinoplanes missouriensis (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 165-172 
    Details
    ISSN: 0887-3585
    Keywords: α/β barrels ; crystal structure ; glucose isomerase ; xylose isomerase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The structure of Xylose isomerase (X.I.) from Actinoplanes missouriensis has been solved to 2.8 Angstroms resolution. Phases were determined from a single Eu3+ derivative and from the noncrystallographic 22 symmetry of the tetrameric molecule. An atomic model was built and subjected to restrained crystallographic refinement. The resulting model is shown to be closely similar to the recently reported X.I.'s structures from three other bacterial sources. Each monomer is found to be composed of an eight-stranded α/β “T.I.M.” barrel forming an N-terminal domain of 328 residues followed by a large loop of 66 residues embracing an adjacent subunit. Analysis of intersubunit packing shows that the X.I. tetramer is an assembly of two tight dimers. The β barrel fits a simple hyperboloid model as other T.I.M. barrels do. The active site, identified as the binding site for the inhibitor xylitol, is located at the carboxyl end of the beta strands in the barrel next to a pair binding site for Eu3+ ions, which are assumed to the sites for the divalent ions involved in catalysis. Active sites in the tetramer are oriented towards the interface between dimmers. It is suggested that subunit interfaces might stabilize the active site region and this might explain the oligomeric nature of the other α/β barrel enzymes.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340040303
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    Formation of heterodimers between wild type and mutant trp aporepressor polypeptides of Escherichia coli (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 173-181 
    Details
    ISSN: 0887-3585
    Keywords: DNA binding protein ; ligand binding ; equilibrium dialysis ; dimer ; stability ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Availability of the three-dimensional structure of the trp repressor of Escherichia coli and a large group of repressor mutants has permitted the identification and analysis of mutants with substitutions of the amino acid residues that from the tryptophan binding pocket. Mutant aporepressors selected for study were overproduced using a multicopy expression plasmid. Equilibrium dialysis with 14C-tryptophan and purified mutant and wild type aporepressors was employed to determine tryptophan binding constants. The results obtained indicate that replacement of theronine 44 by methionine (TM44) or arginine 84 by histidine (RH84) lowers the affinity for tryptophan approximately two-and four-fold, respectively. Replacement of ariginine 54 by histidine (RH84) or glycine 85 by ariginine (GR85) results in complete loss of tryptophan binding activity. Purified mutant and wild type aporepressors were used in vitro heterodimer studies. The trp repressor of E. coli functions as a stable dimer. A large number of trp repressor mutants prduces defective repressors that are transdominant to the wild type repressor in vivo. The transdominance presumably results from the formation of inactive or slightly active heterodimers between the mutant and wild type polypeptide subunits. An in vitro assay was developed to detect and measure heterodimer formation. Heterodimer formation was thermally induced, and heterodimers were separated on nondenaturing polyacrylamide gels. Aporepressors readily formed heterodimer formation upon treatment at 65°C for 3 minutes. Heterodimer formation was significantly retarded by the presence of the corepressor, L-tryptophan. Indole-3-propionic acid, 5-methyl tryptophan, and other analogs of tryptophan, as well as indole, also inhibited heterodimer formation. These results indicate that the presence of the indole moiety in the corepressor binding pocket increases the stability of the dimer.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340040304
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  • 78
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    Interaction of peptide boronic acids with elastase: Circular dichroism studies (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 205-210 
    Details
    ISSN: 0887-3585
    Keywords: slow-binding inhibition ; transition-state analog ; conformational change ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Boronic acid derivatives of good peptide substrate of the serine proteases cause slow-binding inhibition, manifested as biphasic binding (Kettner and Shenvi: J. Biol. Chem. 259:15106-15114, 1984). These inhibitors are thought to act as reaction-intermediate analogs. Three peptides Boronic acids - Ac-Pro-boro-Val-OH, DNS-Ala-Pro-boro-Val-OH, and Ac-Ala-Ala-Pro-boro-Val-OH - were chosen for farultraviolet circular dichroism (CD) studies in order to determine whether the second phase involves a conformational change of pancreatic elastase. The dipeptide is a simple competitive inhibitors (Ki = 0.27 μM) and the latter are slow-binding inhibitors (Ki = 16.4 and 0.25 nM, respectively). Spectral deconvolution and correction for the formation of antiparallel β-sheet by the peptide inhibitors itself indicate that there is no significant change in the secondary structure of the enzyme in the either the initial or final inhibitors complex. A kinetic experiment confirmed that the slow-binding step was not associated with a CD spectral change, and that therefore a protein conformational change was not responsible for the sow binding.
    Additional Material: 2 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340040307
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    Masthead (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988) 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340040401
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  • 80
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    Diffusion-collision model for the folding kinetics of myoglobin (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 211-227 
    Details
    ISSN: 0887-3585
    Keywords: protein-folding ; multiple pathways ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The diffusion-collision model has been used to analyze the folding kinetics of myoglobins. The microdomains, which are the basic units that coalesce during the folding, are identified with the helices and the stabilizing contacts between helices are determined form the native structure. Both association and disassociation reactions are included and a range of stabilization parameters is investigated to determine the variations in overall rate and the relative contributions made by the different intermediates during the folding process. In a comparison of folding to the native state and to the midpoint of the folding transitions. (i.e., 50% native protein at the completion of the reaction) significant differences in the contributing intermediates are found.
    Additional Material: 7 Ill.
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    Test of circular dichroism (CD) methods for crambin and CD-assisted secondary structure prediction of its homologous toxins (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 262-273 
    Details
    ISSN: 0887-3585
    Keywords: hierarchical assignment ; cereal grain ; mistletoe ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Methods that analyze protein circular dichroism (CD) spectra for fractions of secondary structure are evaluated for the plant protein crambin, which has a known high-resolution crystal structure. In addition, a two-step secondary structure prediction scheme is presented and used for the toxins homologous to crambin, shown by others to have secondary structures similar to crambin.The test of CD spectral analysis methods with the protein crambin employed two computer programs and several CD basis sets. Crambin's crystal structure, known to 0.945 Å resolution (Hendrickson, W.A., Teeter, M.M. Nature 290:107-113, 1981), allows accurate evaluation of results. Analysis with the protein spectra basis sets (Provencher, S. W., Glöckner, J. Biochemistry 20:33-37, 1981) as modified (Manavalan, P., Johnson, W. C., Jr. Anal. Biochem. 167:76-85, 1987) agreed most closely with crambin's crystal structure. This method was then applied to the CD spectra of the membrane-active toxins homologous to crambin (α1- and β-purothionin, phoratoxin A and B, an viscotoxin A3 and B).The new program SEQ (pronounced “seek”) was developed to assign the secondary structure along the protein chain in a hierarchical fashion and applied to the plant toxins. The method constrained the secondary structure fractions to those from CD analysis and combined standard statistical methods with amphipathic helix location.Both CD-arrived secondary structure percentages and sequence assignment indicate that the viscotoxins are structurally most similar to crambin. Purothionin's secondary structure was predicted to be fundamentally similar to crambin's with a difference at the start of the first helix. This assignment agreed with Raman and NMR analyses of Purothionin and lends validity to the method presented here. Differences from the NMR in the CD secondary structure fraction analysis for phoratoxin suggest interference in the CD from tryptophan residues.
    Additional Material: 2 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340040405
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    Refined structure of human carbonic anhydrase II at 2.0 Å resolution (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 274-282 
    Details
    ISSN: 0887-3585
    Keywords: crystallography ; refinement ; structure ; carbonic anhydrase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The structure of human erythrocytic carbonic anhydrase II has been refined by constrained and restrained structure-factor least-squares refinement at 2.0 Å resolution. The conventional crystallographic R value is 17.3%. Of 167 solvent molecules associated with the protein, four are buried and stabilize secondary structure elements. The zinc ion is ligated to three histidyl residues and one water molecule in a nearly tetrahedral geometry. In addition to the zinc-bound water, seven more water molecules are identified in the active site. Assuming that Glu-106 is deprotonated at pH 8.5, some of the hydrogen bond donor-acceptor relations in the active site can be assigned and are described here in detail. The Oγ1 atom of Thr-199 donates its proton to the Oε1 atom of Glu-106 and can function as a hydrogen bond acceptor only in additional hydrogen bonds.
    Additional Material: 10 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340040406
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    Stereochemistry of salt-bridge formation in α-helices and β-strands (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 294-295 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340040408
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    Masthead (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988) 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340030101
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  • 85
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    Two trifluoperazine-binding sites on calmodulin predicted from comparative molecular modeling with troponin-C (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 1-17 
    Details
    ISSN: 0887-3585
    Keywords: computer modeling ; trifluoperazine ; conformational change ; calcium binding proteins ; hydrophobic binding interactions ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Among the known regulatory proteins that are conformationally sensitive to the binding of calcium ions, calmodulin and troponin-C have the greatest primary sequence homology. This observation has led to the conclusion that the most accurate predicted molecular model of calmodulin would be based on the X-ray crystallographic coordinates of the highly refined structure of turkey skeletal troponin-C. This paper describes the structure of calmodulin built from such a premise. The resulting molecular model was subjected to conjugate gradient energy minimization to remove unacceptable intramolecular non-bonded contacts. In the analysis of the resulting structure, many features of calmodulin, including the detailed conformation of the Ca2+-binding loops, the amino- and carboxy-terminal hydrophobic patches of the Ca2+-bound form, and the several clusters of acidic residues can be reconciled with much of the previously published solution data. Calmodulin in missing the N-terminal helix characteristic of troponin-C. The deletion of three residues from the central helical linker (denoted D/E in troponin-D) shortens the molecule and changes the orientation of the two domains of calmodulin by 60° relative to those in troponin-C. The molecular model has been used to derive two possible binding sites for the antipsychotic drug trifluoperazine, a potent competitive inhibitor of calmodulin activity.
    Additional Material: 11 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340030102
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  • 86
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    Flexibility of the DNA-binding domains of trp repressor (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 18-31 
    Details
    ISSN: 0887-3585
    Keywords: flexibility ; trp repressor ; DNA-binding domains ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: An orthorhombic crystal form of trp repressor (aporepressor plus L-tryptophan ligand) was solved by molecular replacement, refined to 1.65 Å resolution, and compared to the structure of the repressor in trigonal crystals. Even though these two crystal forms of repressor were grown under identical conditions, the refined structures have distinctly different conformations of the DNA-binding domains. Unlike the repressor/aporepressor structural transition, the conformational shift is not caused by the binding or loss of the L-tryptophan ligand. We conclude that while L-tryptophan binding is essential for forming a specific complex with trp operator DNA, the corepressor ligand does not lock the repressor into a single conformation that is complementary to the operator. This flexibility may be required by the various binding modes proposed for trp repressor in its search for and adherence to its three different operator sites.
    Additional Material: 7 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340030103
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    Ribosome-inhibiting proteins, retroviral reverse transcriptases, and RNase H share common structural elements (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 53-59 
    Details
    ISSN: 0887-3585
    Keywords: ricin ; retroviral integrase ; conserved residues ; homologous sequences ; active site ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Plant ribosome-inhibiting proteins are shown to be homologous at the domain level to RNase H form Escherichia coli and to two regions of the pol gene product of retroviral reverse transcriptases. One of these regions carries the viral integrase or int function, while the other has previously been suggested to contain the viral RNase H exo activity. Several residues conserved among the ribosome inhibitors, E. coli RNase H, and the integrase proteins are seen to occupy a prominent cleft in the tertiary structure of the ribosome inhibitor ricin, suggesting roles in binding or catalysis. It is likely that these homologous sequences represent modern derivatives of an ancient protein-folding unit capable of nucleic acid binding and modification which has been incorporated into a variety of enzyme functions.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340030105
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    Energetics of charge-charge interactions in proteins (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 32-52 
    Details
    ISSN: 0887-3585
    Keywords: alpha-helix ; distance-dependent ; finite-difference method ; Poisson-Boltzmann equation ; protein electrostatics ; rhodanese ; solvent effects ; subtilisin ; Tanford-Kirkwood theory ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Electrostatic interactions between pairs of atoms in proteins are calculated with a model based on the linearized Poisson-Boltzmann equation. The equation is solved accurately by a method that takes into account the detailed shape of the protein. This paper presents applications to several systems. Experimental data for the interaction of ionized residues with an active site histidine in subtilisin BPN' allow the model to be tested, using various assumptions for the electrical properties of the protein and solvent. The electrostatic stabilization of the active site thiolate or rhodanese is analyzed, with attention to the influence of α-helices. Finally, relationships between electrostatic potential and charge-charge distance are reported for large and small globular proteins. The above results are compared with those of simpler electrostatic models, including Coulomb's law with both a distance-dependent dielectric constant (∊ = R) and a fixed dielectric constant (∊ = 2), and Tanford-Kirkwood theory. The primary conclusions are as follows: (1) The Poisson-Boltzmann model agrees with the subtilisin data over a range of ionic strengths; (2) two α-helices generate a large potential in the active site of rhodanese; (3) ∊ = R overestimates weak electrostatic interactions but yields relatively good results for strong ones; (4) Tanford-Kirkwood theory is a useful approximation to detailed solutions of the linearized Poisson-Boltzmann equation in globular proteins; and (5) the modified Tanford-Kirkwood theory overscreens the measured electrostatic interactions in subtilisin.
    Additional Material: 9 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340030104
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    Mechanism of protein folding: I. General considerations and refolding of myoglobin (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 199-207 
    Details
    ISSN: 0887-3585
    Keywords: folding pathway ; hydrophobic interaction ; long-range and long-distance interactions ; secondary structure ; tertiary structure ; module structure ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: To explain the rapidity of the process of protein folding, we cite two aspects of hydrophobic interaction: its long-range nature and the specificity of pairing after the formation of secondary structures. These two factors, when incorporated with the growth-type mechanism, can determine the folding pathway of proteins. This mechanism is applied to myoglobin. Appropriate introduction of side chins of amino acid residues and the heme group attached to His 93 yield a refolded tertiary structure that is in good agreement with the native structure.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340030308
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  • 90
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    Masthead (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988) 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340030401
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  • 91
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    Combined procedure of distance geometry and restrained molecular dynamics techniques for protein structure determination from nuclear magnetic resonance data: Application to the DNA binding domain of lac repressor from Escherichia coli (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 209-218 
    Details
    ISSN: 0887-3585
    Keywords: distance-restrained molecular dynamics ; 2D NOE-spectroscopy ; tertiary structure ; solution conformations ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The technique of two-dimensional nuclear magnetic resonance (2D-NMR) has recently assumed an active role in obtaining information on structures of polypeptides, small proteins, sugars, and DNA fragments in solution. In order to generate spatial structures from the atom-atom distance information obtained by the NMR method, different procedures have been developed. Here we introduce a combined procedure of distance geometry (DG) and molecular dynamics (MD) calculations for generating 3D structures that are consistent with the NMR data set and have reasonable internal energies. We report the application of the combined procedure on the lac repressor DNA binding domain (headpiece) using a set of 169 NOE and 17 “hydrogen bond” distance constraints. Eight of ten structures generated by the distance geometry algorithm were refined within 10 ps MD simulation time to structures with low internal energies that satisfied the distance constraints.Although the combination of DG and MD was designed to combine the good sampling properties of the DG algorithm with an efficient method of lowering the internal energy of the molecule, we found that the MD algorithm contributes significantly to the sampling as well.
    Additional Material: 3 Ill.
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    URL: http://dx.doi.org/10.1002/prot.340030402
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  • 92
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    The aggregation state of spin-labeled melittin in solution and bound to phospholipid membranes: Evidence that membrane-bound melittin is monomeric (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 230-242 
    Details
    ISSN: 0887-3585
    Keywords: melittin ; spin-labelling ; EPR spectroscopy ; membrane-protein interaction ; protein-protein interaction ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Spin-labeled derivatives of the bee venom protein, melittin, were obtained by reacting on the average one of the four amino groups of the protein with succinimidyl-2,2,5,5-tetramethyl-3-pyrroline-1-oxyl-3-carboxylate All 16 statistically possible reaction products with 0, 1, 2, 3 or 4 spin labels per protein were then separated in a single pass with reversed phase high performance liquid chromatography. With the help of trypsin digestion and diode array detection it was possible to assign the primary structure of all 16 eluting fractions. All fractions with only one spin label per protein were purified for electron paramagnetic resonance measurement. The labeling sites cover different regions of the protein: one is at the N-terminus, one at lysine-7, and two are near the C-terminus at lysine-21 and lysine-23, respectively. This set of specifically labeled melittins was used to study the structure and dynamics of melittin in aqueous solutions and when bound to neutral or negatively charged membranes. In aqueous solution a reduction in rotational correlation time and appearance of spin-spin interaction was observed during salt-induced transition from a random coil monomer to a mostly α-helical retramer. Membrane binding to phospholipid bilayers in low or high ionic strength was reflected only in a further decrease in mobility. The absence of any spin interaction in the membrane-bound state suggests that melittin is monomeric under these conditions. All derivatives were able to detect these structural changes, but melittin labeled at the N-terminal amino group was especially valuable. Because of postulated intramolecular hydrogen bonding, this label reflects directly the motion of the entire protein or tetramer. Broadening experiments with chromium oxalate show that all labeled sites are at least partially exposed to the aqueous phase when melittin is bound to membranes. This suggests that an α-helical melittin monomer binds to membranes with its axis parallel to the membrane surface.
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    URL: http://dx.doi.org/10.1002/prot.340030404
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    Preliminary X-ray diffraction studies and biochemical characterization of the antitumor protein mitomalcin indicate close similarity to neocarzinostatin (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 252-255 
    Details
    ISSN: 0887-3585
    Keywords: streptomyces malayensis ; antitumor antibiotic ; holoprotein antibiotic ; crystallization of mitomalcin ; amino acid composition ; partial amino acid sequence ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The antitumor antibiotic protein mitomalcin, from the microorganism Streptomyces malayensis, has been purified to apparent homogenity and crystallized. The crystals belong to space group P212121 and have the following cell parameters : a=27.2 Å, b=34.1 Å, c=101.7 Å, and alpha;=β=γ=90°. These crystal properties are extremely similar to crystals of the antitumor protein neocarzinostatin (11.7 kilodaltons [kDa]) from Streptomyces carzinostaticus in spite of differing pH conditions for crystallizing the two proteins and an apparent difference in molecular weight is similar to that of neocarzinostatin. An amino acid composition analysis of mitomalcin indicates that some differences may exist between the two molecules, but a preliminary amino acid sequence analysis of the first 37 residues found no difference in the N-terminal region of the molecule.
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    URL: http://dx.doi.org/10.1002/prot.340030406
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  • 94
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    Unusual zymogen-processing properties of a mutated form of prochymosin (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 256-261 
    Details
    ISSN: 0887-3585
    Keywords: zymogen activation ; protein engineering ; autoproteolytic processing ; protein structure/function ; renaturation ; rDNA expression ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Site-specific mutagenesis of the gene encoding bovine prochymosin was used to produce a mutated zymogen in which seven contiguous amino acids of the N-terminal propeptide had been deleted and an eighth residue had been substituted. This altered region spans the normal site of autocatalytic proteolysis that occurs at the same time as (enzymatic) activation of prochymosin at acidic pH. Activation of the mutated zymogen at pH 4.5 was extremely slow, and cleavage occurred at an unusual Ser-Lys bond in the prochymosin incubated at pH 2 generated the usual pseudochymosin by cleavage of the normal Phe-Leu bond, but at a rate severalfold slower than the authentic zymogen. These results indicate that even after deletion of seven of 42 amino acids of the propeptide the mutant protein could still assume a prochymosin (zymogen) structure, although these changes did result in striking differences in acid-catalyzed activation and processing reactions at one but not the other of the two processing sites of prochymosin.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340030407
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  • 95
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    Electronic Resource
    Masthead (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988) 
    Details
    ISSN: 0887-3585
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340040101
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  • 96
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    Microfolding: Conformational probability map for the alanine dipeptide in water from molecular dynamics simulations (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 3 (1988), S. 262-265 
    Details
    ISSN: 0887-3585
    Keywords: protein conformation ; conformational equilibria ; free energy stimulation ; protein folding ; thermodynamic perturbation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A direct attack on the protein-folding problem has been initiated with the free energy perturbation methods of molecular dynamics. The complete conformational probability map for the alanine dipeptide is presented. This work uses the SPC model for the explicit hydration of the dipeptide. Free energy differences for the four observed minima (β, αR, αL, C7ax) are given, and the free energy barriers between minima are outlined.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340030408
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  • 97
    Electronic Resource
    Electronic Resource
    Homology of lysosomal enzymes and related proteins: Prediction of posttranslational modification sites including phosphorylation of mannose and potential epitopic and substrate binding sites in the α- and β-subunits of hexosaminidases, α-glucosidase, and rabbit and human isomaltase (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 182-189 
    Details
    ISSN: 0887-3585
    Keywords: consensus sequences ; secondary structure ; mannose 6-phosphate ; substrate specificity ; proteolytic processing ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Recently developed computer programs, including secondary structure and epitopic site predictions, have been used to align lysosomal proteins for maximum homology, based on conservative interchanges, and the aligned sequences have been searched for potential sites for posttranslational modification, glycosyaltion, and binding and catalysis of substrate. The homology and prediction of the posttranslational modification of the α- and β-subunits of hexosaminidase is in good agreement with previous observations, and an explanation of the differing substrate specificities of the two subunits is advanced. We shows that the striking homology between α-glucosidase and isomaltase is reflected in that apparent conversation of the active site in both enzymes. Nonhomologous regions have been examined in detail in a search for binding sited for glycogen and maltose, and two such sites have been tentatively identified. A highly redundant consensus sequence for the Phosphorylation of mannose in lysosomal proteins, YXX(Y, W, or F), is suggested.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340040305
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  • 98
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    Electronic Resource
    Comparative molecular model building of two serine proteinases from cytotoxic T lymphocytes (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 190-204 
    Details
    ISSN: 0887-3585
    Keywords: computer graphics ; energy minimization ; proteolytic enzymes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Two genes that are expressed when precursor cytotoxic T lymphocytes are transformed to T killer cells have been cloned and sequenced. The derived amino acid sequences, coding for the cytotoxic cell protease 1 (CCP1) and Hannuka factor (HF) are highly homologous to members of the serine proteinase family. Comparative molecular model building using the known three-dimensional structures and the derived amino acid sequences of the lymphocyte enzymes has been provided useful information, especially in predicting the conformations of the substrate binding sites. In applying this modelling procedure, we used the X-ray structures of four serine proteinase to provide a structurally based sequences alignment: α-chymotrypsin (CHT), bovine trypsin (BT), Streptomyces griseus trypsin (SGT), and rat must cell protease 2 (RMCP2). The root mean square differences in α-carbon atom positions among these four structures when compared in a pairwise fashions range form 0.79 to 0.97 Å for structurally equivalent residues. Te sequences of the two lymphocyte enzymes were then aligned to these proteinase using chemical criteria and the superimposed X-ray structures as guides. The alignment showed that the sequence of CCP1 was most similar to RMCP2, whereas HF has regions of homology with both RMCP2 and BT. RmCP2 and BT as templates for HF, the molecular models were constructed. Intermolecular steric clashes that resulted from replacement of amino acid chains of the templates by the aligned residues of CCP1 and HF were relieved by adjustment of the side chain conformational angels in an interactive computer graphics device. This process was followed by energy minimization of the enzyme model to optimize the stereochemical geometry and to relieve any remaining unacceptably close nonbonded contacts. The resulting model of CCP1 has an arginie residue at position 226 in the specificity pocket, thereby predicting a substrate preference for P1 aspartate or glutamate residues. The model also predicts favorable binding for a small hydrophobic residue at the P2 position of the substrate. The primary specificity pocket of HF resembles that of BT and therefore predicts a lysine or arginine preference for the P1 residue. The arginine at position 99 in the model of HF suggests a preference for aspartate or glutamate side chains in the P2 positions of the substrate. Both CCP1 and HF have a free cysteine in the segment of the polypeptide 88 to 93. Models of the dimeric form of these enzymes can be constructed by forming disulfide bridges between the suitably oriented monomers, ie., Cys88-Cys88′ for CCP1 and Cys93-Cys93′ for HF.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340040306
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  • 99
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    Electronic Resource
    Helix lap-joints as ion-binding sites: DNA-binding motifs and Ca-binding “EF hands” are related by charge and sequence reversal (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 229-239 
    Details
    ISSN: 0887-3585
    Keywords: repressor proteins ; helix dipole ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The DNA-binding helix pairs in gene repressor and activator were compared with other approximately perpendicular pairs of adjacent helices in the known protein structures. Two other examples of closely matching conformations were found in cytochrome c peroxidase (residues 153-174) and in ribosomal L7/L12 protein (residues 68-89). Another group of such offset “lap-joints” are the Ca-binding “EF hand” structures, which bind a positive rather than a negative ligand. The EF hands turn out to match the DNA-binding motifs quite well (outside of the loop) if their sequence direction is reversed. This conformation is thus not as unusual as had been thought, but may have a more generalized role in ion binding and occasionally occur in a purely structural role.
    Additional Material: 12 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340040402
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  • 100
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    Structural alignment and analysis of two distantly related proteins: Aplysia limacina myoglobin and sea lamprey globin (1988)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 4 (1988), S. 240-250 
    Details
    ISSN: 0887-3585
    Keywords: protein structure ; X-Ray crystal structure ; homology ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Two new globin structures have recently been determined at high resolution: the globin from the mollusc Aplysia limacina at 1.6 A resolution and a new refinement of the structure from sea lamprey. Two amino acid sequences of these homologous molecules have only 30% residue identity in an optimal alignment. We discuss some of the problems arising in the alignment of Aplysia globin with other globins of known structure, a challenging problem because of the distant relationship. Four independent approaches were applied to the alignment of the Aplysia and lamprey globins, including those based on individual sequence comparisons, structural analysis, and the relatively new method of templates or fingerprints derived for an entire family of proteins. We also compare these two new structures with what is already known about the globin family. A detailed description of the two structures shows that the two molecules contain the main structural features common to all the globins so far studied with several minor but interesting hitherto unobserved variations.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340040403
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