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  • Artikel: DFG Deutsche Nationallizenzen  (56)
  • Cell & Developmental Biology  (38)
  • Biochemistry and Biotechnology  (18)
  • 1
    ISSN: 0887-3585
    Schlagwort(e): structure-based drug design ; ligand-protein recognition ; binding landscapes ; docking funnels ; stochastic search ; Chemistry ; Biochemistry and Biotechnology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Medizin
    Notizen: Energy landscapes of molecular recognition are explored by performing “semi-rigid” docking of FK-506 and rapamycin with the Fukisawa binding protein (FKBP-12), and flexible docking simulations of the Ro-31-8959 and AG-1284 inhibitors with HIV-1 protease by a genetic algorithm. The requirements of a molecular recognition model to meet thermodynamic and kinetic criteria of ligand-protein docking simultaneously are investigated using a family of simple molecular recognition energy functions. The critical factor that determines the success rate in predicting the structure of ligand-protein complexes is found to be the roughness of the binding energy landscape, in accordance with a minimal frustration principle. The results suggest that further progress in structure prediction of ligand-protein complexes can be achieved by designing molecular recognition energy functions that generate binding landscapes with reduced frustration. © 1996 Wiley-Liss, Inc.
    Zusätzliches Material: 10 Ill.
    Materialart: Digitale Medien
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  • 2
    Digitale Medien
    Digitale Medien
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 29 (1997), S. 59-67 
    ISSN: 0887-3585
    Schlagwort(e): blind trials ; CASP2 ; homology modelling ; protein structure prediction ; Chemistry ; Biochemistry and Biotechnology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Medizin
    Notizen: The CASP blind trials (Critical Assessment of techniques for protein Structure Prediction) assess the accuracy of protein prediction that includes evaluation of comparative model building of protein structures. Comparative models of four proteins (T0001, T0003, T0017, and T0028) for CASP2 (held during 1996) were constructed using computer algorithms combined with visual inspection. Essentially the main-chain modelling involves construction of the target structure from rigid-body segments of homologues and loop fragments extracted from homologous and nonredundant databases. Side-chains were initially constructed by inheritance from the parent or from a rotamer library. Side-chain conformations were then refined using a novel mean field approach that includes solvation. Comparison of the models with the subsequently released X-ray structures identified the successes and limitations of our approach. The most problematic area is the quality of the sequence alignments between parent(s) and target. In this respect the overinterpretation of the conserved features within homologous families can be misleading. Several features of our approach have a positive effect on the accuracy of the models. For T0003, inspection correctly identified that a lower sequence identity parent provides the best framework for this model. Loop selection worked well where a homologous protein fragment was used, but that the use of nonredundant fragment library remains problematic for hinge movements and displacements in secondary structure elements relative to the parent. Side-chain refinement improved residue conformations relative to the initial model. Use of limited energy minimization improved the stereochemical quality of the model without increasing the RMS deviation. This study has identified methods that are effective and areas requiring further attention to improve model building by comparison. Proteins, Suppl. 1:59-67, 1997. © 1998 Wiley-Liss, Inc.
    Zusätzliches Material: 5 Ill.
    Materialart: Digitale Medien
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  • 3
    Digitale Medien
    Digitale Medien
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 25 (1993), S. 87-104 
    ISSN: 0886-1544
    Schlagwort(e): polymerization ; solation ; gelation ; α-actinin ; gelsolin ; calcium ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie , Medizin
    Notizen: We describe a cellular automaton model of the actin cytoskeleton. The model incorporates spatial and temporal behavior at the macomolecular level and is relevant to the viscous nonequilibrium conditions suspected to occur in vivo. The model include cation and nucleotide binding to actin monomers, actin nucleation and polymerization into filaments, coss-linking with α-actinin, monomer sequestration with pfilin, filament severing, capping and nucleation with gelsolin, binding of profilin and gelsolin to membrane-bound phosphatidylinositide biphosphate (PIP2), and regulation of coss-linking and severing by changing calcium levels. We derive (1) equations for the molecular trnslation and rotation probabilities required for the cellular automaton simulation in terms of molecular size, shape, cytoplasmic viscosity, and temperature; and (2) equations for the binding probabilities of adjacent molecules in terms of experimentally determined reaction rate constants. The model accurately captures the known characteristics of actin polymerization and subsequent ATP hydrolysis under different cation and nucleotide conditions. An examination of gelation and sol-gel transitions resulting from calcium regulation of α-actinin and gelsolin predicts an inhomogeneous distribution of bound α-actinin and F-actin. The double-bound α-actinin (both ends bound to F-actin) is tightly bunched, while single-bound α-actinin is moderately bunched and unbound α-actinin is homogeneously distributed. The spatial organization of the α-actinin is quantified using estimates of fractal dimension. The simulation results also suggest that actin/α-actinin gels may shift from an isotropic to an amorphous phase after shortening of filaments. The gel-sol transition of the model shows excellent agreement with the present theory of polymer gels. The close correspondence of the model's predictions with previous experimental and theoretical results suggests that the model may be pertinent to better understanding the spatial and temporal properties of complex cytoskeletal processes. © 1993 Wiley-Liss, Inc.
    Zusätzliches Material: 9 Ill.
    Materialart: Digitale Medien
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  • 4
    Digitale Medien
    Digitale Medien
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 28 (1997), S. 313-324 
    ISSN: 0887-3585
    Schlagwort(e): molecular docking ; receptor-specific binding ; binding energy landscapes ; structural water molecules ; minimal frustration principle ; Chemistry ; Biochemistry and Biotechnology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Medizin
    Notizen: Mean field analysis of FKBP12 complexes with FK506 and rapamycin has been performed by using structures obtained from molecular docking simulations on a simple, yet robust molecular recognition energy landscape. When crystallographic water molecules are included in the simulations as an extension of the FKBP12 protein surface, there is an appreciable stability gap between the energy of the native FKBP12-FK506 complex and energies of conformations with the “native-like” binding mode. By contrast, the energy spectrum of the FKBP12-rapamycin complex is dense regardless of the presence of the water molecules. The stability gap in the FKBP12-FK506 system is determined by two critical water molecules from the effector region that participate in a network of specific hydrogen bond interactions. This interaction pattern protects the integrity and precision of the composite ligand-protein effector surface in the binary FKBP12-FK506 complex and is preserved in the crystal structure of the FKBP12-FK506-calcineurin ternary complex. These features of the binding energy landscapes provide useful insights into specific and nonspecific aspects of FK506 and rapamycin recognition. Proteins 28:313-324, 1997. © 1997 Wiley-Liss, Inc.
    Zusätzliches Material: 9 Ill.
    Materialart: Digitale Medien
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  • 5
    Digitale Medien
    Digitale Medien
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 28 (1997), S. 421-433 
    ISSN: 0887-3585
    Schlagwort(e): molecular recognition ; binding energy landscapes ; recognition nucleus ; structural harmony ; minimal frustration principle ; Chemistry ; Biochemistry and Biotechnology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Medizin
    Notizen: Computational structure prediction of streptavidin-peptide complexes for known recognition sequences and a number of random di-, tri-, and tetrapeptides has been conducted, and mechanisms of peptide recognition with streptavidin have been investigated by a new computational protocol. The structural consensus criterion, which is computed from multiple docking simulations and measures the accessibility of the dominant binding mode, identifies recognition motifs from a set of random peptide sequences, whereas energetic analysis is less discriminatory. The predicted conformations of recognition tripeptide and tetrapeptide sequences are also in structural harmony and composed of peptide fragments that are individually unfrustrated in their bound conformation, resulting in a minimally frustrated energy landscape for recognition peptides. Proteins 28:421-433, 1997. © 1997 Wiley-Liss, Inc.
    Zusätzliches Material: 8 Ill.
    Materialart: Digitale Medien
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  • 6
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 26 (1990), S. 248-252 
    ISSN: 1040-452X
    Schlagwort(e): Microfilaments ; Pinocytosis ; Amino acids ; Defolliculation ; Pigment ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie
    Notizen: Defolliculated fully grown oocytes of Xenopus laevis were treated with cytochalasin D (10 μg/ml) and their protein synthesis was studied by labelling with S-35 methionine. This treatment brought about an alteration in pigment pattern as well as a reduction in amino acid uptake by the oocytes. However, the radioactive amino acid taken by cytochalasin-treated oocytes was incorporated into protein in the same proportion as in untreated oocytes. These results suggested that subcortical pigment distribution and amino acid uptake in fully grown oocytes were microfilament-dependent processes, whereas protein synthesis in the oocyte was not.
    Zusätzliches Material: 5 Ill.
    Materialart: Digitale Medien
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  • 7
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 36 (1993), S. 7-15 
    ISSN: 1040-452X
    Schlagwort(e): Intercellular coupling ; Connexin32 ; Connexin43 ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie
    Notizen: The connexins constitute a family of proteins that make up the intercellular membrane channels of gap junctions. We had previously reported the presence of two members of this protein family, connexins 32 and 43, in mouse one-cell zygotes (Barron et al., Dev Genet 10:318-323, 1989; Valdimarsson et al., Mol Reprod Dev 30:18-26, 1991), implying that both must be present in the mature oocyte and could be involved in mediating the intercellular coupling that occurs between the oocyte and cumulus granulosa during oogenesis. In the present report we provide evidence for this, based on an analysis of the cumulus-oocyte complex (COC) using reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry with a confocal microscope. Transcripts of both connexin32 (Cx32) and connexin43 (Cx43) were detected by RT-PCR in both components of the COC. Cx32 mRNA in the oocyte declined precipitously following human chorionic gonadotropin (hCG) stimulation of pregnant mare serum gonadotropin (PMSG)-primed ovaries, whereas there was no obvious change in Cx43 mRNA. Peptide-specific antibodies against both connexins provided diffuse cytoplasmic staining of oocytes as well as some punctate staining near the oocyte surface, which could not be unequivocally resolved as cumulus-oocyte gap junctions. However, the two antibodies did provide clear evidence of Cx32 and Cx43 in gap junction-like structures between cumulus cells. We could find no evidence of the incorporation of the oocyte's store of Cx32 into gap junctions during postfertilization development. These findings make it very likely that intercellular coupling within the COC involves at least three types of gap junction channels (32-32, 32-43, 43-43), only one of which (43-43) is retained by the preimplantation embryo. © 1993 Wiley-Liss, Inc.
    Zusätzliches Material: 4 Ill.
    Materialart: Digitale Medien
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  • 8
    Digitale Medien
    Digitale Medien
    Chicester [u.a.] : Wiley-Blackwell
    Journal of Molecular Recognition 3 (1990), S. 215-219 
    ISSN: 0952-3499
    Schlagwort(e): Chemistry ; Biochemistry and Biotechnology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Medizin
    Notizen: We have previously shown that citrate synthase binds to an intrinsic protein of the mitochondrial inner membrane (D'Souza and Srere, 1983). In this paper we present evidence that this citrate synthase binding protein is the citrate transporter. We have used citrate synthase 1 mutants of Saccharomyces cerevesiae and transformants containing citrate synthase inactivated by site-directed mutagenesis to study the effect of the CS1 protein upon mitochondrial function (Kispal and Srere). In the present study citrate uptake and oxidation were measured during state 3 conditions (presence of 200 μM ADP) in the mitochondria of several strains of Saccharomyces cerevesiae: a parental strain containing wild-type mitochondrial citrate synthase (CS1) and strains derived from a CS1 deficient strain in which the CS1 gene was disrupted by insertation of the LEU2 gene. These strains were generated from the CS1- cells by transformation with vectors encoding site-specific mutants of CS1 possessing very low levels of enzymatic activity. One such strain in this study was subsequently found to have undergone reversion to produce a strain which had activity very similar to wild type. Positive correlation between citrate uptake and the rate of citrate oxidation was found, suggesting coupling of the two processes. Both mitochondrial citrate uptake and oxidation were decreased in the mutant lacking any form of CS1 protein. Reintroduction of mutagenized CS1 into yeast causes an enhancement in the rate of state 3 oxygen consumption and of citrate uptake. The observed respiratory increase produces a state 3 oxygen consumption rate which is 1.5- to 17-fold greater than the rate of reintroduced mutagenized CS1 activity; conversely, wild-type CS1 enzyme levels are about 30-fold greater than the respiratory rate in wild-type cells. Further evidence for uncoupling of the process of citrate utilization from CS1 activity is also seen in the revertant strain which has lower respiratory rate than the wild-type strain. This difference is not due to decreased concentration of the citrate carrier of those cells with lower respiratory rates, a change in equilibrium for citrate across the mitochondrial membrane, or to decreased citrate saturability of such mitochondria. These results, supported by the interaction observed during column chromatography between matrix-immobilized pig citrate synthase and the protein from yeast mitochondrial membrane preparations which exhibits citrate transport activity, strongly suggest physical interaction between CS1 and the mitochondrial citrate transport protein.
    Zusätzliches Material: 3 Ill.
    Materialart: Digitale Medien
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  • 9
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 30 (1991), S. 18-26 
    ISSN: 1040-452X
    Schlagwort(e): Gap junction protein ; Gene expression ; Compaction ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie
    Notizen: De novo assembly of gap junctions begins during compaction in the eight-cell stage of mouse development, and intercellular coupling mediated by gap junctions appears to be required for maintenance of the compacted state. We have begun to explore the expression of the family of genes encoding the connexins, the proteins that form the gap junction channels. We recently reported that a protein with antigenic and size similarity with connexin32, the rat liver gap junction protein, is inherited as an oogenetic product by the mouse zygote, but its gene appears not to be transcribed prior to implantation (Barron et al., Dev Genet 10:318-323, 1989). Here we report that another member of this gene family, connexin43, is transcribed by the embryonic genome from shortly after the time of genomic activation. As revealed by Northern blotting, connexin43 mRNA is absent from ovulated oocytes, becomes detectable in the 4-cell stage, and accumulates steadily thereafter to reach a maximum in blastocysts. In contrast, no transcripts of connexin26 could be detected in any preimplantation stage. A protein with antigenic and size similarity with connexin43 from rat heart was found by Western blotting to accumulate from the four-cell stage onward. Immunofluorescence analysis with embryo whole mounts was used to demonstrate that this protein is incorporated into punctate interblastomeric foci during compaction, consistent with its assembly into gap junction plaques. We conclude that connexin43 is one member of the connexin gene family whose zygotic expression is critical for preimplantation morphogenesis.
    Zusätzliches Material: 6 Ill.
    Materialart: Digitale Medien
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  • 10
    Digitale Medien
    Digitale Medien
    Chicester [u.a.] : Wiley-Blackwell
    Journal of Molecular Recognition 1 (1988), S. 19-24 
    ISSN: 0952-3499
    Schlagwort(e): Chemistry ; Biochemistry and Biotechnology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Medizin
    Notizen: We have investigated a possible molecular basis for mitochondrial cristae formation. Proteoliposomes containing electron transport proteins, cytochrome oxidase, or complex III in their proper orientation bind to pig heart mitoplasts but not pig heart mitochondria. Using Leydig tumor cells, we have confirmed earlier reports that chloramphenicol causes a diminution in cristae content and a change in its characteristic lamellar form. We show that the proteoliposomes containing cytochrome oxidase or complex III in the proper orientation bind to mitoplasts from Leydig tumor cells but do not bind as well to mitoplasts from chloramphenicol-treated Leydig tumor cells. These experiments provide a possible mechanism to explain cristae formation.
    Zusätzliches Material: 3 Ill.
    Materialart: Digitale Medien
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