Molecular basis for substrate specificity of protein-tyrosine phosphatase 1B

Mauro Sarmiento, Yu Zhao, Steven J. Gordon, Zhong-Yin Zhang

Research output: Contribution to journalArticle

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Abstract

Protein-tyrosine phosphatases can exhibit stringent substrate specificity in vivo, although the molecular basis for this is not well understood. The three-dimensional structure of the catalytically inactive protein-tyrosine phosphate 1B (PTP1B)/C215S complexed with an optimal substrate, DADEpYL-NH2, reveals specific interactions between amino acid residues in the substrate and PTP1B. The goal of this work is to rigorously evaluate the functional significance of Tyr46, Arg47, Asp48, Phe182, and Gln262 in substrate binding and catalysis, using site-directed mutagenesis. Combined with structural information, kinetic analysis of the wild type and mutant PTP1B using p-nitrophenyl phosphate and phosphotyrosine- containing peptides has yielded further insight into PTP1B residues, which recognize general features, as well as specific properties, in peptide substrates. In addition, the kinetic results suggest roles of these residues in E-P hydrolysis, which are not obvious from the structure of PTP1B/peptide complex. Thus, Tyr46 and Asp48 recognize common features of peptide substrates and are important for peptide substrate binding and/or E-P formation. Arg47 acts as a determinant of substrate specificity and is responsible for the modest preference of PTP1B for acidic residues NH2- terminal to phosphotyrosine. Phe182 and the invariant Gln262 are not only important for substrate binding and/or E-P formation but also important for the E-P hydrolysis step.

Original languageEnglish (US)
Pages (from-to)26368-26374
Number of pages7
JournalJournal of Biological Chemistry
Volume273
Issue number41
DOIs
StatePublished - Oct 9 1998

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Non-Receptor Type 1 Protein Tyrosine Phosphatase
Substrate Specificity
Tyrosine
Phosphates
Substrates
Peptides
Phosphotyrosine
Proteins
Hydrolysis
Protein Tyrosine Phosphatases
Mutant Proteins
Site-Directed Mutagenesis
Catalysis
Mutagenesis
Kinetics
Amino Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Molecular basis for substrate specificity of protein-tyrosine phosphatase 1B. / Sarmiento, Mauro; Zhao, Yu; Gordon, Steven J.; Zhang, Zhong Yin.

In: Journal of Biological Chemistry, Vol. 273, No. 41, 09.10.1998, p. 26368-26374.

Research output: Contribution to journalArticle

Sarmiento, Mauro ; Zhao, Yu ; Gordon, Steven J. ; Zhang, Zhong Yin. / Molecular basis for substrate specificity of protein-tyrosine phosphatase 1B. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 41. pp. 26368-26374.
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