Abstract
Rho Kinase I (ROCK I) is a serine/threonine kinase that is involved in diverse cellular signaling. To further understand the physiological role of ROCK I and to identify and develop potent and selective inhibitors of ROCK I, we have overexpressed and purified a constitutively active dimeric human ROCK I (3-543) kinase domain using the Sf9-baculovirus expression system. In addition, using a limited proteolysis technique, we have identified a minimal functional subdomain of ROCK I that can be used in crystallization studies. The availability of multimilligram amounts of purified and well characterized functional human ROCK I kinase domains will be useful in screening and structural studies.
MeSH terms
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1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / analogs & derivatives
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1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / pharmacology
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Amides / pharmacology
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Amino Acid Sequence
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Animals
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Humans
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Inhibitory Concentration 50
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Intracellular Signaling Peptides and Proteins
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Molecular Sequence Data
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Molecular Weight
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Protein Serine-Threonine Kinases / antagonists & inhibitors
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Protein Serine-Threonine Kinases / biosynthesis*
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Protein Serine-Threonine Kinases / isolation & purification
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Protein Serine-Threonine Kinases / metabolism
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Protein Structure, Tertiary
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Pyridines / pharmacology
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Spodoptera
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Staurosporine / pharmacology
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rho-Associated Kinases
Substances
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Amides
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Intracellular Signaling Peptides and Proteins
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Pyridines
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Y 27632
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1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
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Protein Serine-Threonine Kinases
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rho-Associated Kinases
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Staurosporine
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fasudil