The Experts below are selected from a list of 23202 Experts worldwide ranked by ideXlab platform
Stephen Oroszlan - One of the best experts on this subject based on the ideXlab platform.
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Purification of HIV‐1 wild‐type protease and characterization of proteolytically inactive HIV‐1 protease mutants by Pepstatin A affinity chromatography
FEBS letters, 1991Co-Authors: Ewald M. Wondrak, John M. Louis, Peter T. Mora, Stephen OroszlanAbstract:Recombinant wild-type protease of human immunodeficiency virus, type [(HIV-1) expressed in E. coli was purified by Pepstatin A affinity chromatography. An 88-fold purification was achieved giving a protease preparation with a specific enzymatic activity of approximately 3700 pmol/min/μg. Two proteolytically inactive HIV-1 mutant proteases (Arg-87 → Lys; Asn-88 → Glu) were found to bind to Pepstatin A agarose, and they were purified as the wild-type protease. A third mutant protease (Arg-87 → Glu) was apparently unable to bind to Pepstatin A under similar conditions. Binding to Pepstatin A indicates the binding ability of the substrate binding site and the ability to form dimers. These features may be used to purify and to characterize other mutated HIV-1 proteases.
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purification of hiv 1 wild type protease and characterization of proteolytically inactive hiv 1 protease mutants by Pepstatin a affinity chromatography
FEBS Letters, 1991Co-Authors: Ewald M. Wondrak, John M. Louis, Peter T. Mora, Stephen OroszlanAbstract:Recombinant wild-type protease of human immunodeficiency virus, type [(HIV-1) expressed in E. coli was purified by Pepstatin A affinity chromatography. An 88-fold purification was achieved giving a protease preparation with a specific enzymatic activity of approximately 3700 pmol/min/μg. Two proteolytically inactive HIV-1 mutant proteases (Arg-87 → Lys; Asn-88 → Glu) were found to bind to Pepstatin A agarose, and they were purified as the wild-type protease. A third mutant protease (Arg-87 → Glu) was apparently unable to bind to Pepstatin A under similar conditions. Binding to Pepstatin A indicates the binding ability of the substrate binding site and the ability to form dimers. These features may be used to purify and to characterize other mutated HIV-1 proteases.
Ewald M. Wondrak - One of the best experts on this subject based on the ideXlab platform.
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Purification of HIV‐1 wild‐type protease and characterization of proteolytically inactive HIV‐1 protease mutants by Pepstatin A affinity chromatography
FEBS letters, 1991Co-Authors: Ewald M. Wondrak, John M. Louis, Peter T. Mora, Stephen OroszlanAbstract:Recombinant wild-type protease of human immunodeficiency virus, type [(HIV-1) expressed in E. coli was purified by Pepstatin A affinity chromatography. An 88-fold purification was achieved giving a protease preparation with a specific enzymatic activity of approximately 3700 pmol/min/μg. Two proteolytically inactive HIV-1 mutant proteases (Arg-87 → Lys; Asn-88 → Glu) were found to bind to Pepstatin A agarose, and they were purified as the wild-type protease. A third mutant protease (Arg-87 → Glu) was apparently unable to bind to Pepstatin A under similar conditions. Binding to Pepstatin A indicates the binding ability of the substrate binding site and the ability to form dimers. These features may be used to purify and to characterize other mutated HIV-1 proteases.
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purification of hiv 1 wild type protease and characterization of proteolytically inactive hiv 1 protease mutants by Pepstatin a affinity chromatography
FEBS Letters, 1991Co-Authors: Ewald M. Wondrak, John M. Louis, Peter T. Mora, Stephen OroszlanAbstract:Recombinant wild-type protease of human immunodeficiency virus, type [(HIV-1) expressed in E. coli was purified by Pepstatin A affinity chromatography. An 88-fold purification was achieved giving a protease preparation with a specific enzymatic activity of approximately 3700 pmol/min/μg. Two proteolytically inactive HIV-1 mutant proteases (Arg-87 → Lys; Asn-88 → Glu) were found to bind to Pepstatin A agarose, and they were purified as the wild-type protease. A third mutant protease (Arg-87 → Glu) was apparently unable to bind to Pepstatin A under similar conditions. Binding to Pepstatin A indicates the binding ability of the substrate binding site and the ability to form dimers. These features may be used to purify and to characterize other mutated HIV-1 proteases.
József Tőzsér - One of the best experts on this subject based on the ideXlab platform.
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inhibition of xmrv and hiv 1 proteases by Pepstatin a and acetyl Pepstatin
FEBS Journal, 2012Co-Authors: Krisztina Matúz, János András Mótyán, Alexander Wlodawer, József TőzsérAbstract:The kinetic properties of two classical inhibitors of aspartic proteases (PRs), Pepstatin A and acetyl-Pepstatin, were compared in their interactions with HIV-1 and xenotropic murine leukemia virus related virus (XMRV) PRs. Both compounds are substantially weaker inhibitors of XMRV PR than of HIV-1 PR. Previous kinetic and structural studies characterized HIV-1 PR-acetyl-Pepstatin and XMRV PR-Pepstatin A complexes and suggested dramatically different binding modes. Interaction energies were calculated for the possible binding modes and suggested a strong preference for the one-inhibitor binding mode for HIV-1 PR-acetyl-Pepstatin and the two-inhibitor binding mode for XMRV PR-Pepstatin A interactions. Comparison of the molecular models suggested that in the case of XMRV PR the relatively unfavorable interactions at S3' and the favorable interactions at S4 and S4' sites with the statine residues may shift the ground state binding towards the two-inhibitor binding mode, whereas the single molecule ground state binding of statines to the HIV-1 PR appear to be more favorable. The preferred single molecular binding to HIV-1 PR allows the formation of the transition state complex, represented by substantially better binding constants. Intriguingly, the crystal structure of the complex of acetyl-Pepstatin with XMRV PR has shown a mixed type of binding: the unusual binding mode of two molecules of the inhibitor to the enzyme, in a mode very similar to the previously determined complex with Pepstatin A, together with the classical binding mode found for HIV-1 PR. The structure is thus in good agreement with the very similar interaction energies calculated for the two types of binding.
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Inhibition of XMRV and HIV‐1 proteases by Pepstatin A and acetyl‐Pepstatin
The FEBS journal, 2012Co-Authors: Krisztina Matúz, János András Mótyán, Alexander Wlodawer, József TőzsérAbstract:The kinetic properties of two classical inhibitors of aspartic proteases (PRs), Pepstatin A and acetyl-Pepstatin, were compared in their interactions with HIV-1 and xenotropic murine leukemia virus related virus (XMRV) PRs. Both compounds are substantially weaker inhibitors of XMRV PR than of HIV-1 PR. Previous kinetic and structural studies characterized HIV-1 PR-acetyl-Pepstatin and XMRV PR-Pepstatin A complexes and suggested dramatically different binding modes. Interaction energies were calculated for the possible binding modes and suggested a strong preference for the one-inhibitor binding mode for HIV-1 PR-acetyl-Pepstatin and the two-inhibitor binding mode for XMRV PR-Pepstatin A interactions. Comparison of the molecular models suggested that in the case of XMRV PR the relatively unfavorable interactions at S3' and the favorable interactions at S4 and S4' sites with the statine residues may shift the ground state binding towards the two-inhibitor binding mode, whereas the single molecule ground state binding of statines to the HIV-1 PR appear to be more favorable. The preferred single molecular binding to HIV-1 PR allows the formation of the transition state complex, represented by substantially better binding constants. Intriguingly, the crystal structure of the complex of acetyl-Pepstatin with XMRV PR has shown a mixed type of binding: the unusual binding mode of two molecules of the inhibitor to the enzyme, in a mode very similar to the previously determined complex with Pepstatin A, together with the classical binding mode found for HIV-1 PR. The structure is thus in good agreement with the very similar interaction energies calculated for the two types of binding.
Peter T. Mora - One of the best experts on this subject based on the ideXlab platform.
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Purification of HIV‐1 wild‐type protease and characterization of proteolytically inactive HIV‐1 protease mutants by Pepstatin A affinity chromatography
FEBS letters, 1991Co-Authors: Ewald M. Wondrak, John M. Louis, Peter T. Mora, Stephen OroszlanAbstract:Recombinant wild-type protease of human immunodeficiency virus, type [(HIV-1) expressed in E. coli was purified by Pepstatin A affinity chromatography. An 88-fold purification was achieved giving a protease preparation with a specific enzymatic activity of approximately 3700 pmol/min/μg. Two proteolytically inactive HIV-1 mutant proteases (Arg-87 → Lys; Asn-88 → Glu) were found to bind to Pepstatin A agarose, and they were purified as the wild-type protease. A third mutant protease (Arg-87 → Glu) was apparently unable to bind to Pepstatin A under similar conditions. Binding to Pepstatin A indicates the binding ability of the substrate binding site and the ability to form dimers. These features may be used to purify and to characterize other mutated HIV-1 proteases.
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purification of hiv 1 wild type protease and characterization of proteolytically inactive hiv 1 protease mutants by Pepstatin a affinity chromatography
FEBS Letters, 1991Co-Authors: Ewald M. Wondrak, John M. Louis, Peter T. Mora, Stephen OroszlanAbstract:Recombinant wild-type protease of human immunodeficiency virus, type [(HIV-1) expressed in E. coli was purified by Pepstatin A affinity chromatography. An 88-fold purification was achieved giving a protease preparation with a specific enzymatic activity of approximately 3700 pmol/min/μg. Two proteolytically inactive HIV-1 mutant proteases (Arg-87 → Lys; Asn-88 → Glu) were found to bind to Pepstatin A agarose, and they were purified as the wild-type protease. A third mutant protease (Arg-87 → Glu) was apparently unable to bind to Pepstatin A under similar conditions. Binding to Pepstatin A indicates the binding ability of the substrate binding site and the ability to form dimers. These features may be used to purify and to characterize other mutated HIV-1 proteases.
John M. Louis - One of the best experts on this subject based on the ideXlab platform.
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Purification of HIV‐1 wild‐type protease and characterization of proteolytically inactive HIV‐1 protease mutants by Pepstatin A affinity chromatography
FEBS letters, 1991Co-Authors: Ewald M. Wondrak, John M. Louis, Peter T. Mora, Stephen OroszlanAbstract:Recombinant wild-type protease of human immunodeficiency virus, type [(HIV-1) expressed in E. coli was purified by Pepstatin A affinity chromatography. An 88-fold purification was achieved giving a protease preparation with a specific enzymatic activity of approximately 3700 pmol/min/μg. Two proteolytically inactive HIV-1 mutant proteases (Arg-87 → Lys; Asn-88 → Glu) were found to bind to Pepstatin A agarose, and they were purified as the wild-type protease. A third mutant protease (Arg-87 → Glu) was apparently unable to bind to Pepstatin A under similar conditions. Binding to Pepstatin A indicates the binding ability of the substrate binding site and the ability to form dimers. These features may be used to purify and to characterize other mutated HIV-1 proteases.
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purification of hiv 1 wild type protease and characterization of proteolytically inactive hiv 1 protease mutants by Pepstatin a affinity chromatography
FEBS Letters, 1991Co-Authors: Ewald M. Wondrak, John M. Louis, Peter T. Mora, Stephen OroszlanAbstract:Recombinant wild-type protease of human immunodeficiency virus, type [(HIV-1) expressed in E. coli was purified by Pepstatin A affinity chromatography. An 88-fold purification was achieved giving a protease preparation with a specific enzymatic activity of approximately 3700 pmol/min/μg. Two proteolytically inactive HIV-1 mutant proteases (Arg-87 → Lys; Asn-88 → Glu) were found to bind to Pepstatin A agarose, and they were purified as the wild-type protease. A third mutant protease (Arg-87 → Glu) was apparently unable to bind to Pepstatin A under similar conditions. Binding to Pepstatin A indicates the binding ability of the substrate binding site and the ability to form dimers. These features may be used to purify and to characterize other mutated HIV-1 proteases.
