Mechanism of inhibition of protein-tyrosine phosphatases by vanadate and pervanadate

Gregory Huyer, Susana Liu, John Kelly, Jason Moffat, Paul Payette, Brian Kennedy, George Tsaprailis, Michael J. Gresser, Chidambaram Ramachandran

Research output: Contribution to journalArticlepeer-review

741 Scopus citations


Vanadate and pervanadate (the complexes of vanadate with hydrogen peroxide) are two commonly used general protein-tyrosine phosphatase (PTP) inhibitors. These compounds also have insulin-mimetic properties, an observation that has generated a great deal of interest and study. Since a careful kinetic study of the two inhibitors has been lacking, we sought to analyze their mechanisms of inhibition. Our results show that vanadate is a competitive inhibitor for the protein-tyrosine phosphatase PTP1B, with a K(i) of 0.38 ± 0.02 μM. EDTA, which is known to chelate vanadate, causes an immediate and complete reversal of the inhibition due to vanadate when added to an enzyme assay. Pervanadate, by contrast, inhibits by irreversibly oxidizing the catalytic cysteine of PTP1B, as determined by mass spectrometry. Reducing agents such as dithiothreitol that are used in PTP assays to keep the catalytic cysteine reduced and active were found to convert pervanadate rapidly to vanadate. Under certain conditions, slow time- dependent inactivation by vanadate was observed; since catalose blocked this inactivation, it was ascribed to in situ generation of hydrogen peroxide and subsequent formation of pervanodate. Implications for the use of these compounds as inhibitors and rationalization for some of their in vivo effects are considered.

Original languageEnglish (US)
Pages (from-to)843-851
Number of pages9
JournalJournal of Biological Chemistry
Issue number2
StatePublished - 1997
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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