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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of biomedical science 4 (1997), S. 235-243 
    ISSN: 1423-0127
    Keywords: Kistrin ; Disintegrin ; Integrin ; Extracellular matrix ; Metastasis ; Bone marrow stromal cell
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Rhodostomin from venom ofAgkistrodon rhodostoma (also calledCalloselasma rhodostoma) contains 68 amino acid residues including 6 pairs of disulfide bonds and an arginine-glycine-aspartic acid (RGD) sequence at positions 49–51. It has been known as one of the strongest antagonists to platelet aggregation among the family termed disintegrin. In this review paper, in addition to introducing the characteristics of disintegrin and its related molecules, the advantages of using recombinant DNA technology to produce rhodostomin are described. The recombinant rhodostomin has been demonstrated to facilitate cell adhesion via interaction between the RGD motif of rhodostomin and integrins on the cell surface. This property allowed us to use the recombinant rhodostomin as an extracellular matrix to study cell adhesion and to distinguish attachment efficiency between two melanoma cell lines B16-F1 and B16-F10, the former is a low metastasis cell while the latter is a high metastasis cell. Furthermore, by using the recombinant rhodostomin as a substrate, osteoprogenitor-like cells are able to be selected and enriched within 3 days from rat bone marrow which contains a heterogeneous cell population. Finally, we show that the recombinant rhodostomin can be immobilized on beads and which serve as an affinity column to dissect cell-surface protein(s) binding to the RGD motif of rhodostomin.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Journal of biomedical science 6 (1999), S. 64-70 
    ISSN: 1423-0127
    Keywords: Bacterial display ; TraT lipoprotein ; Outer membrane protein ; Rhodostomin ; Hepatitis B virus pre-S1 antigen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract ThetraT gene is one of the F factor transfer genes and encodes an outer membrane protein which is involved in interactions between anEscherichia coli and its surroundings. This protein was altered so as to permit the expression of foreign proteins on the outer membrane ofE. coli in this study. A 729-bp DNA fragment, including the leader and entire structural gene sequence oftraT, was amplified and obtained by PCR. This sequence was then subcloned downstream of thetac promoter of pDR540, resulting in a TraT expression vector, pT2. Here, we report that the expression of TraT protein, fused either with a partial pre-S antigen of hepatitis B virus (60 and 98 amino acids, respectively) or with the snake venom rhodostomin (72 amino acids), was successfully achieved on the outer membrane ofE. coli, using the pT2 plasmid. This result was demonstrated using dot blot and immunofluorescence analysis. This finding supports the notion that the pT2 plasmid can be used as anE. coli display system. This system can detect a foreign peptide of about 100 amino acid residues in length on the bacterial surface.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Journal of biomedical science 7 (2000), S. 42-50 
    ISSN: 1423-0127
    Keywords: Endocytosis, receptor-mediated ; Bacterial display ; TraT lipoprotein ; Rhodostomin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Previously, we developed a TraT display system to express snake venom rhodostomin (RHO), a disintegrin, on the external surface ofEscherichia coli [J Biomed Sci 6:64–70;1999]. To show a new potential use of the TraT display system, we employed a biotin labeling technique coupled with SDS-PAGE and flow cytometry analyses to further demonstrate and confirm the expression of TraT-RHO on theE. coli surface. We also showed that the expression of TraT-RHO on the cell surface not only facilitated the bacteria adhesion to BHK-21 cells but also induced bacterial internalization into BHK-21 cells. This feature allowed us to enrich the TraT-RHO expression bacteria about 10,000-fold starting with a mixture of TraT-RHO bacteria with β-galactosidase-positive bacteria in a ratio of 102:107 through four cycles of BHK-21 cell endocytosis and replating of engulfed bacteria on agar plates. We therefore suggest that the TraT display system can be applied to select out bacteria expressing a specific peptide sequence from a large population of display library through the process of receptor-mediated endocytosis and reamplification cycles.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1572-994X
    Keywords: herpes simplex virus (HSV) ; latency associated transcript (LAT) ; ribozyme ; reactivation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The latency associated transcript (LAT) of herpes simplex virus (HSV) appears to exist as an RNA molecule only. This phenomenon is consistent with the concept of functioning at an RNA level, and several lines of evidence suggest that the LAT may be a ribozyme. This provides an insight into understanding the role of LAT during HSV reactivation.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 0886-1544
    Keywords: pigment organelle ; xanthophore ; microtubule ; F-actin ; intermediate filament ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In goldfish xanthophores, the formation of pigment aggregate requires: (1) that a pigment organelle (carotenoid droplet) protein p57 be in the unphosphorylated state; (2) that self-association of pigment organelles occur in a microtubule-independent manner; and (3) that pigment organelles via p57 associate with microtubules. In the fully aggregated state, the pigment organelles are completely stationary. Pigment dispersion is initiated by activation of a cAMP-dependent protein kinase, which phosphorylates p57 and allows pigment dispersion via an active process dependent on F-actin and a cytosolic factor. This factor is not an ATPase, and its function is unknown. However, its abundance in different tissues parallels secretory activity of the tissues, suggesting a similarity between secretion and pigment dispersion in xanthophores. The identity of the motor for pigment dispersion is unclear. Experimental results show that pigment organelles isolated from cells with dispersed pigment have associated actin and ATPase activity comparable to myosin ATPase. This ATPase is probably an organelle protein of relative molecular mass ∼72,000, and unlikely to be an ion pump. Isolated pigment organelles without associated actin have 5× lower ATPase activity. Whether this organelle ATPase is the motor for pigment dispersion is under investigation. The process of pigment aggregation is poorly understood, with conflicting results for and against the involvement of intermediate filaments.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 13 (1989), S. 9-20 
    ISSN: 0886-1544
    Keywords: carotenoid droplet ; intermediate filament ; microfilament ; microtubule ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The cytoskeleton of goldfish xanthophores contains an abundance of unique dense structures (400 nm in diameter) that are absent in goldfish nonpigment cells and are probably remnants of pterinosomes. No major difference in protein composition between xanthophores and nonpigment cells (without these structures) was found that could account for these structures. In xanthophores, these structures are foci of radiating filaments. The addition or withdrawal of ACTH causes a radical rearrangement of the xanthophore Cytoskeleton accompanying redistribution of carotenoid droplets, namely, the virtual exclusion of these dense bodies with associated filaments from the space occupied by the carotenoid droplet aggregate vs. a relatively even cytoplasmic distribution of these structures when the carotenoid droplets are dispersed. These changes in cytoskeletal morphology are not accompanied by any major changes in the protein or phosphoprotein composition of the cytoskeleton.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-0878
    Keywords: Filopodia ; Microfilaments ; Cellular processes ; Xanthophores ; Pigment cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Treatment of cultured goldfish xanthophores by hormone (ACTH) or c-AMP induces not only pigment dispersion, but subsequent outgrowth of processes, and pigment translocation into these processes. These latter effects are shown to proceed as follows: First the edge of the cytoplasmic lamellae takes on a scalloped contour with numerous protrusions. These presumably serve as nucleation centers where short microfilament bundles are assembled, Later, the microfilament bundles elongate (“grow”), often resulting in an extension of the protrusions to become filopodia while the proximal end of the microfilaments penetrates into the thicker portion of the cellular process which now houses the pigment, i.e., the carotenoid droplets. Carotenoid droplets appear to migrate along the microfilament bundles, or cytoplasmic channels associated with them, into the filopodia. Finally, some of the filopodia become broader, thicker and laden with carotenoid droplets and are then recognized by light microscopy as pigmented cellular processes. The microfilaments have been shown to be actin filaments by their thickness, the size of their subunits, and decoration by heavy meromyosin. Evidence is presented which suggests that the growth of these actin filaments may come about by recruitment from short F-actin strands found in random orientation in adjacent areas.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-0878
    Keywords: Endoplasmic reticulum ; Actin filaments ; Xanthophores ; Pigment cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The hormone-induced pigment dispersion in primary cultures of xanthophores of goldfish (Carassius auratus L.) has been shown to involve the dispersion of not only carotenoid droplets but also of smooth endoplasmic reticulum. The dispersion of these organelles is inhibited by cytochalasin B and is accompanied by thinning of the cell body, thickening of the processes, and also overall changes in cellular morphology (process extension) under certain conditions. Electron microscopic examination of heavy meromyosin treated glycerinated xanthophores in scales revealed the presence of actin filaments in these cells.
    Type of Medium: Electronic Resource
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