Abstract
Rho-kinase binds to a small GTPase Rho in a GTP-dependent manner and regulates many cytoskeletal events in the cell. The minimum region of bovine Rho-kinase sufficient for Rho-binding was expressed as a fusion protein with glutathione S-transferase. After removal of the glutathione S-transferase, thin plate crystals were obtained. The selenomethionine-substituted protein was introduced and crystallized, as was the native protein. The crystals of the Rho-binding domain of Rho-kinase belong to the space group C2, with unit-cell parameters a = 148.0 (2), b = 26.1 (1), c = 39.6 (1) A, beta = 90.3 (1) degrees. The crystals diffract to a resolution beyond 1.5 A.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Binding Sites
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Cattle
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Crystallization
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Escherichia coli / genetics
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Glutathione Transferase / chemistry
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Glutathione Transferase / genetics
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Glutathione Transferase / isolation & purification
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Intracellular Signaling Peptides and Proteins
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Protein Serine-Threonine Kinases / chemistry*
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Protein Serine-Threonine Kinases / genetics
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Protein Serine-Threonine Kinases / isolation & purification
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Protein Structure, Tertiary
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / isolation & purification
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rho GTP-Binding Proteins / metabolism
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rho-Associated Kinases
Substances
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Intracellular Signaling Peptides and Proteins
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Recombinant Fusion Proteins
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Glutathione Transferase
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Protein Serine-Threonine Kinases
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rho-Associated Kinases
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rho GTP-Binding Proteins