Protein tyrosine phosphatases regulate signal transduction pathways involving tyrosine phosphorylation1 and have been implicated in the development of cancer, diabetes, rheumatoid arthritis and hypertension2. Increasing evidence suggests that the cellular redox state is involved in regulating tyrosine phosphatase activity through the reversible oxidization of the catalytic cysteine to sulphenic acid (Cys-SOH)3,4,5,6. But how further oxidation to the irreversible sulphinic (Cys-SO2H) and sulphonic (Cys-SO3H) forms is prevented remains unclear. Here we report the crystal structures of the regulatory sulphenic and irreversible sulphinic and sulphonic acids of protein tyrosine phosphatase 1B (PTP1B), an important enzyme in the negative regulation of the insulin receptor7,8 and a therapeutic target in type II diabetes and obesity9. We also identify a sulphenyl-amide species that is formed through oxidation of its catalytic cysteine. Formation of the sulphenyl-amide causes large changes in the PTP1B active site, which are reversible by reduction with the cellular reducing agent glutathione. The sulphenyl-amide is a protective intermediate in the oxidative inhibition of PTP1B. In addition, it may facilitate reactivation of PTP1B by biological thiols and signal a unique state of the protein.
This is a preview of subscription content, access via your institution
Open Access articles citing this article.
Nature Communications Open Access 26 June 2020
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
Neel, B. G. & Tonks, N. K. Protein tyrosine phosphatases in signal transduction. Curr. Opin. Cell Biol. 9, 193–204 (1997)
Zhang, Z. Y. Protein tyrosine phosphatases: structure and function, substrate specificity, and inhibitor development. Annu. Rev. Pharmacol. Toxicol. 42, 209–234 (2002)
Denu, J. M. & Tanner, K. G. Specific and reversible inactivation of protein tyrosine phosphatases by hydrogen peroxide: evidence for a sulphenic acid intermediate and implications for redox regulation. Biochemistry 37, 5633–5642 (1998)
Denu, J. M. & Dixon, J. E. Protein tyrosine phosphatases: mechanisms of catalysis and regulation. Curr. Opin. Chem. Biol. 2, 633–641 (1998)
Meng, T. C., Fukada, T. & Tonks, N. K. Reversible oxidation and inactivation of protein tyrosine phosphatases in vivo. Mol. Cell 9, 387–399 (2002)
Lee, S. R., Kwon, K. S., Kim, S. R. & Rhee, S. G. Reversible inactivation of protein-tyrosine phosphatase 1B in A431 cells stimulated with epidermal growth factor. J. Biol. Chem. 273, 15366–15372 (1998)
Elchebly, M. et al. Increased insulin sensitivity and obesity resistance in mice lacking the protein tyrosine phosphatase-1B gene. Science 283, 1544–1548 (1999)
Klaman, L. D. et al. Increased energy expenditure, decreased adiposity, and tissue-specific insulin sensitivity in protein-tyrosine phosphatase 1B-deficient mice. Mol. Cell. Biol. 20, 5479–5489 (2000)
Goldstein, B. J. Protein-tyrosine phosphatase 1B (PTP1B): a novel therapeutic target for type 2 diabetes mellitus, obesity and related states of insulin resistance. Curr. Drug. Targets Immune Endocr. Metabol. Disord. 1, 265–275 (2001)
Barford, D. Protein phosphatases. Curr. Opin. Struct. Biol. 5, 728–734 (1995)
Pannifer, A. D., Flint, A. J., Tonks, N. K. & Barford, D. Visualization of the cysteinyl-phosphate intermediate of a protein-tyrosine phosphatase by X-ray crystallography. J. Biol. Chem. 273, 10454–10462 (1998)
Barford, D., Flint, A. J. & Tonks, N. K. Crystal structure of human protein tyrosine phosphatase 1B. Science 263, 1397–1404 (1994)
Jia, Z., Barford, D., Flint, A. J. & Tonks, N. K. Structural basis for phosphotyrosine peptide recognition by protein tyrosine phosphatase 1B. Science 268, 1754–1758 (1995)
Lohse, D. L., Denu, J. M., Santoro, N. & Dixon, J. E. Roles of aspartic acid-181 and serine-222 in intermediate formation and hydrolysis of the mammalian protein-tyrosine-phosphatase PTP1. Biochemistry 36, 4568–4575 (1997)
Sarmiento, M., Zhao, Y., Gordon, S. J. & Zhang, Z. Y. Molecular basis for substrate specificity of protein-tyrosine phosphatase 1B. J. Biol. Chem. 273, 26368–26374 (1998)
Scapin, G., Patel, S., Patel, V., Kennedy, B. & Asante-Appiah, E. The structure of apo protein-tyrosine phosphatase 1B C215S mutant: more than just an S → O change. Protein Sci. 10, 1596–1605 (2001)
Mahadev, K., Zilbering, A., Zhu, L. & Goldstein, B. J. Insulin-stimulated hydrogen peroxide reversibly inhibits protein-tyrosine phosphatase 1b in vivo and enhances the early insulin action cascade. J. Biol. Chem. 276, 21938–21942 (2001)
Claiborne, A. et al. Protein-sulfenic acids: diverse roles for an unlikely player in enzyme catalysis and redox regulation. Biochemistry 38, 15407–15416 (1999)
Claiborne, A., Miller, H., Parsonage, D. & Ross, R. P. Protein-sulfenic acid stabilization and function in enzyme catalysis and gene regulation. FASEB J. 7, 1483–1490 (1993)
Barford, D., Kelly, J., Flint, A. J. & Tonks, N. K. Purification and crystallization of the catalytic domain of human protein tyrosine phosphatase 1B expressed in Escherichia coli. J. Mol. Biol. 239, 726–730 (1994)
Pflugrath, J. W. The finer things in X-ray diffraction data collection. Acta Crystallogr. D 55, 1718–1725 (1999)
Collaborative Computational Project No. 4. The CCP4 suite: programs for protein crystallography. Acta Crystallogr. D 50, 760–763 (1994)
Hartshorn, M. J. A visualisation aid for structure-based drug design. J. Comput. Aid. Mol. Des. (in the press)
We thank N. Wallis and G. Williams for discussions; O. Callaghan for compounds; I. Tickle for assistance with crystallographic software; and D. Barford for access to his data before submission and for discussions.
The authors declare that they have no competing financial interests.
About this article
Cite this article
van Montfort, R., Congreve, M., Tisi, D. et al. Oxidation state of the active-site cysteine in protein tyrosine phosphatase 1B. Nature 423, 773–777 (2003). https://doi.org/10.1038/nature01681
This article is cited by
The role of protein tyrosine phosphatase 1B (PTP1B) in the pathogenesis of type 2 diabetes mellitus and its complications
Journal of Physiology and Biochemistry (2022)
Nature Chemical Biology (2020)
Nature Communications (2020)
Nature Chemical Biology (2018)
Nature Chemical Biology (2018)