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Mutagenesis of the catalytic subunit of rabbit muscle protein phosphatase-1
Authors:Zhang  Zhongjian  Zhao  Sumin  Deans-Zirattu  Stephen  Bai  Ge  Lee  Ernest Y C
Institution:(1) Department of Biochemistry & Molecular Biology, University of Miami School of Medicine, P.O. Box 016129, 33101 Miami, FL, USA
Abstract:We have generated site-directed mutants of the catalytic subunit of rabbit muscle ppase-1. Since it is known that ppase-1 and ppase-2A are highly susceptible to inactivation by sulfhydryl reagents, we have mutagenized the six cysteine residues conserved between these two enzymes to serines. The six mutants were purified to near homogeneity by affinity chromatography on inhibitor-2-Sepharose and characterized. All six exhibited enzymatic activity. These results indicate that the catalytic mechanism of ppase-1 is different from that of the protein tyrosine phosphatases which involve a cysteinyl phosphate intermediate.
Keywords:protein phosphatase-1  mutagenesis of protein phosphatase-1  cysteine residues  phosphatase-1
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