The second messenger hydrogen peroxide is required for optimal activation of numerous signal transduction pathways, particularly those mediated by protein tyrosine kinases1,2,3,4,5,6. One mechanism by which hydrogen peroxide regulates cellular processes is the transient inhibition of protein tyrosine phosphatases through the reversible oxidization of their catalytic cysteine, which suppresses protein dephosphorylation7,8,9. Here we describe a structural analysis of the redox-dependent regulation of protein tyrosine phosphatase 1B (PTP1B), which is reversibly inhibited by oxidation after cells are stimulated with insulin8 and epidermal growth factor9. The sulphenic acid intermediate produced in response to PTP1B oxidation is rapidly converted into a previously unknown sulphenyl-amide species, in which the sulphur atom of the catalytic cysteine is covalently linked to the main chain nitrogen of an adjacent residue. Oxidation of PTP1B to the sulphenyl-amide form is accompanied by large conformational changes in the catalytic site that inhibit substrate binding. We propose that this unusual protein modification both protects the active-site cysteine residue of PTP1B from irreversible oxidation to sulphonic acid and permits redox regulation of the enzyme by promoting its reversible reduction by thiols.
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Finkel, T. Oxygen radicals and signalling. Curr. Opin. Cell Biol. 10, 248–253 (1998)
Finkel, T. Oxidants painting the cysteine chapel: redox regulation of PTPs. Dev. Cell 2, 251–259 (2002)
Rhee, S. G., Bae, Y. S., Lee, S. R. & Kwon, J. Hydrogen peroxide: a key messenger that modulates protein phosphorylation through cysteine oxidation. Sci. STKE 53 (2000); available at 〈http://www.stke.org/cgi/content/full/OC_sigtrans;2000/53/pe1〉
Sundaresan, M., Yu, Z. X., Ferrans, V. J., Irani, K. & Finkel, T. Requirement for generation of H2O2 for platelet-derived growth factor signal transduction. Science 270, 296–299 (1995)
Rao, G. N. Hydrogen peroxide induces complex formation of SHC–Grb2–SOS with receptor tyrosine kinase and activates Ras and extracellular signal-regulated protein kinases group of mitogen-activated protein kinases. Oncogene 13, 713–719 (1996)
Bae, Y. S. et al. Epidermal growth factor (EGF)-induced generation of hydrogen peroxide. Role in EGF receptor-mediated tyrosine phosphorylation. J. Biol. Chem. 272, 217–221 (1997)
Knebel, A., Rahmsdorf, H. J., Ullrich, A. & Herrlich, P. Dephosphorylation of receptor tyrosine kinases as target of regulation by radiation, oxidants or alkylating agents. EMBO J. 15, 5314–5325 (1996)
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)
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)
Barford, D., Das, A. K. & Egloff, M.-P. The structure and mechanism of protein phosphatases: insights into catalysis and regulation. Annu. Rev. Biophys. Biomol. Struct. 27, 133–164 (1998)
Denu, J. M. & Tanner, K. G. Specific and reversible inactivation of protein tyrosine phosphatases by hydrogen peroxide: evidence for a sulfenic acid intermediate and implications for redox regulation. Biochemistry 37, 5633–5642 (1998)
Collaborative Computational Project No. 4. The CCP4 suite: programs for protein crystallography. Acta Crystallogr. D 50, 760–763 (1994)
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)
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)
Flint, A. J., Tiganis, T., Barford, D. & Tonks, N. K. Development of ‘substrate-trapping’ mutants to identify physiological substrates of protein tyrosine phosphatases. Proc. Natl Acad. Sci. USA 94, 1680–1685 (1997)
Elchelbly, 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)
Haj, F. G., Markova, B., Klaman, L. D., Bohmer, F. D. & Neel, B. G. Regulation of receptor tyrosine kinase signaling by protein tyrosine phosphatase-1B. J. Biol. Chem. 278, 739–744 (2003)
Caselli, A. et al. The inactivation mechanism of low molecular weight phosphotyrosine-protein phosphatase by H2O2 . J. Biol. Chem. 273, 32554–32560 (1998)
Lee, S. R. et al. Reversible inactivation of the tumour suppressor PTEN by H2O2 . J. Biol. Chem. 277, 20336–20342 (2002)
Barrett, W. C. et al. Regulation of PTP1B via glutathionylation of the active site cysteine 215. Biochemistry 38, 6699–6705 (1999)
Kim, S. O. et al. OxyR: a molecular code for redox-related signaling. Cell 109, 383–396 (2002)
Blanchetot, C., Tertoolen, L. G. & den Hertog, J. Regulation of receptor protein-tyrosine phosphatase alpha by oxidative stress. EMBO J. 15, 493–503 (2002)
Otwinowski, Z. & Minor, W. Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 307–326 (1997)
Brunger, A. T. et al. Crystallography & NMR system: a new software suite for macromolecular structure determination. Acta Crystallogr. D 54, 905–921 (1998)
We thank H. Jhoti for communicating data before submission. This work was funded by grants from Cancer Research UK (D.B.) and the NIH (N.K.T.).
The authors declare that they have no competing financial interests.
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Salmeen, A., Andersen, J., Myers, M. et al. Redox regulation of protein tyrosine phosphatase 1B involves a sulphenyl-amide intermediate. Nature 423, 769–773 (2003). https://doi.org/10.1038/nature01680
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