Dissecting the catalytic mechanism of protein-tyrosine phosphatases

Zhong-Yin Zhang, Yuan Wang, Jack E. Dixon

Research output: Contribution to journalArticle

240 Citations (Scopus)

Abstract

Protein-tyrosine phosphatases (PTPases) contain an evolutionarily conserved segment of 250 amino acids referred to as the PTPase catalytic domain. The recombinant PTPase domain from Yersinia enterocolitica enhances the rate of hydrolysis of p-nitrophenyl phosphate, a phosphate monoester, by approximately 1011 over the non-enzyme-catalyzed rate by water. Specific amino acid residues responsible for the catalytic rate acceleration have been examined by site-directed mutagenesis. Our results suggest that Asp-356 (D356) and Glu-290 (E290) are the general acid and the general base catalysts responsible for Yersinia PTPase-catalyzed phosphate ester hydrolysis. The PTPase with both E290Q and D356N mutations shows no pH dependence for catalysis but displays a rate enhancement of 2.6 x 106, compared to the noncatalyzed hydrolysis of p-nitrophenyl phosphate by water. This rate enhancement probably occurs via transition-state stabilization. Our results suggest that all PTPases use a common mechanism that depends upon formation of a thiol-phosphate intermediate and general acid-general base catalysis.

Original languageEnglish (US)
Pages (from-to)1624-1627
Number of pages4
JournalProceedings of the National Academy of Sciences of the United States of America
Volume91
Issue number5
StatePublished - Mar 1 1994
Externally publishedYes

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Protein Tyrosine Phosphatases
Hydrolysis
Phosphates
Catalysis
Amino Acids
Yersinia
Yersinia enterocolitica
Acids
Water
Site-Directed Mutagenesis
Sulfhydryl Compounds
Catalytic Domain
Esters
Mutation

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Dissecting the catalytic mechanism of protein-tyrosine phosphatases. / Zhang, Zhong-Yin; Wang, Yuan; Dixon, Jack E.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 91, No. 5, 01.03.1994, p. 1624-1627.

Research output: Contribution to journalArticle

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