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Redox regulation of protein tyrosine phosphatase 1B involves a sulphenyl-amide intermediate


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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Figure 1: Oxidation of PTP1B results in formation of a sulphenyl-amide bond between Cys 215 and Ser 216.
Figure 2: Conformational changes accompanying the oxidation of PTP1B.
Figure 3: Pulse-chase analysis of oxidation of PTP1B in solution.
Figure 4: Effects of oxidation on substrate binding and phosphorylation of PTP1B.


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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.).

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Correspondence to David Barford.

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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).

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