Identincation of a novel aryl phosphate-binding site in ptp1b

implications for inhibitor design

Zhong Yin Zhan, Yoram A. Puius, Yu Zhao, Michael Sullivan, David S. Lawrence, Steven C. Almo

Research output: Contribution to journalArticle

Abstract

The structure of the catalytically inactive mutant (C215S) of the human protein-tyrosine phosphatase IB (PTP1B) has been solved to high resolution in two complexes. In the first, crystals were grown in the presence of bis(-4nra-phosphophenyl)methane (BPPM), a synthetic high-affinity low molecular weight non-peptidic substrate (Km=l6 |iM), and the structure was refined to an R-factor of 18.2% at 1.9 A resolution. In the second, crystals were grown in a saturating concentration of phosphotyrosine (pTyr), and the structure was refined to an R-factor of 18.1 % at 1.85 A. Difference Fourier maps showed that BPPM binds FTP IB in two mutually exclusive modes, one in which it occupies the canonical pTyr-binding site (the active site), and another in which a phosphophenyl moiety interacts with a set of residues not previously observed to bind aryl phosphates. The identification of a second pTyr molecule at the same site in the PTPlB/C215S-pTyr complex confirms that these residues constitute a low-affinity non-catalytic aryl phosphatebinding site. Although the biological relevance of the second aryl phosphate binding site is unclear, its identification provides a new paradigm for the design of tight-binding, highly specific inhibitors for PTP1B.

Original languageEnglish (US)
JournalFASEB Journal
Volume11
Issue number9
StatePublished - 1997

Fingerprint

Phosphotyrosine
crystals
R388
binding sites
Phosphates
Binding Sites
phosphates
protein-tyrosine-phosphatase
active sites
methane
molecular weight
mutants
Crystals
Protein Tyrosine Phosphatases
Methane
Catalytic Domain
Molecular Weight
Molecular weight
Molecules
Substrates

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Identincation of a novel aryl phosphate-binding site in ptp1b : implications for inhibitor design. / Zhan, Zhong Yin; Puius, Yoram A.; Zhao, Yu; Sullivan, Michael; Lawrence, David S.; Almo, Steven C.

In: FASEB Journal, Vol. 11, No. 9, 1997.

Research output: Contribution to journalArticle

Zhan, Zhong Yin ; Puius, Yoram A. ; Zhao, Yu ; Sullivan, Michael ; Lawrence, David S. ; Almo, Steven C. / Identincation of a novel aryl phosphate-binding site in ptp1b : implications for inhibitor design. In: FASEB Journal. 1997 ; Vol. 11, No. 9.
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AU - Zhan, Zhong Yin

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AU - Lawrence, David S.

AU - Almo, Steven C.

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