Oxidation state of the active-site cysteine in protein tyrosine phosphatase 1B

Protein tyrosine phosphatases regulate signal transduction pathways involving tyrosine phosphorylation and have been implicated in the development of cancer, diabetes, rheumatoid arthritis and hypertension. 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). 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 receptor and a therapeutic target in type II diabetes and obesity. 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.

[1]  L. Lally.  The CCP 4 Suite — Computer programs for protein crystallography , 1998 .

[2]  Z. Otwinowski,et al.  Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[3]  T. Finkel.  Oxygen radicals and signaling. , 1998, Current opinion in cell biology.

[4]  Y. Zhao,et al.  Molecular Basis for Substrate Specificity of Protein-tyrosine Phosphatase 1B* , 1998, The Journal of Biological Chemistry.

[5]  V. Ferrans,et al.  Requirement for Generation of H2O2 for Platelet-Derived Growth Factor Signal Transduction , 1995, Science.

[6]  D. Barford,et al.  The structure and mechanism of protein phosphatases: insights into catalysis and regulation. , 1998, Annual review of biophysics and biomolecular structure.

[7]  Joanne I. Yeh,et al.  Protein-sulfenic acids: diverse roles for an unlikely player in enzyme catalysis and redox regulation. , 1999, Biochemistry.

[8]  A. Ullrich,et al.  Dephosphorylation of receptor tyrosine kinases as target of regulation by radiation, oxidants or alkylating agents. , 1996, The EMBO journal.

[9]  R J Read,et al.  Crystallography & NMR system: A new software suite for macromolecular structure determination. , 1998, Acta crystallographica. Section D, Biological crystallography.

[10]  J. Denu,et al.  Specific and reversible inactivation of protein tyrosine phosphatases by hydrogen peroxide: evidence for a sulfenic acid intermediate and implications for redox regulation. , 1998, Biochemistry.

[11]  D. Barford.  Protein Phosphatases , 2004, Topics in Current Genetics.

[12]  J. Dixon,et al.  Roles of aspartic acid-181 and serine-222 in intermediate formation and hydrolysis of the mammalian protein-tyrosine-phosphatase PTP1. , 1997, Biochemistry.

[13]  G. Rao.  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. , 1996, Oncogene.

[14]  L. Zhu,et al.  Insulin-stimulated Hydrogen Peroxide Reversibly Inhibits Protein-tyrosine Phosphatase 1B in Vivo and Enhances the Early Insulin Action Cascade* , 2001, The Journal of Biological Chemistry.

[15]  Zhon-Yin Zhang,et al.  Protein tyrosine phosphatases: structure and function, substrate specificity, and inhibitor development. , 2002, Annual review of pharmacology and toxicology.

[16]  B. Kennedy,et al.  Increased insulin sensitivity and obesity resistance in mice lacking the protein tyrosine phosphatase-1B gene. , 1999, Science.

[17]  Toshiyuki Fukada,et al.  Reversible oxidation and inactivation of protein tyrosine phosphatases in vivo. , 2002, Molecular cell.

[18]  F. Haj,et al.  Regulation of Receptor Tyrosine Kinase Signaling by Protein Tyrosine Phosphatase-1B* , 2003, The Journal of Biological Chemistry.

[19]  Michael J. Hartshorn,et al.  AstexViewerTM †: a visualisation aid for structure-based drug design , 2002, J. Comput. Aided Mol. Des..

[20]  E. Asante-Appiah,et al.  The structure of apo protein‐tyrosine phosphatase 1B C215S mutant: More than just an S → O change , 2001, Protein science : a publication of the Protein Society.

[21]  S. Rhee,et al.  Reversible Inactivation of Protein-tyrosine Phosphatase 1B in A431 Cells Stimulated with Epidermal Growth Factor* , 1998, The Journal of Biological Chemistry.

[22]  T. Onytiganis.  Development of ''substrate-trapping'' mutants to identify physiological substrates of protein tyrosine phosphatases , 1997 .

[23]  D. Parsonage,et al.  Protein‐sulfenic acid stabilization and function in enzyme catalysis and gene regulation , 1993, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[24]  Young-Bum Kim,et al.  Increased Energy Expenditure, Decreased Adiposity, and Tissue-Specific Insulin Sensitivity in Protein-Tyrosine Phosphatase 1B-Deficient Mice , 2000, Molecular and Cellular Biology.

[25]  T. Finkel,et al.  Oxidants painting the cysteine chapel: redox regulation of PTPs. , 2002, Developmental cell.

[26]  D. Barford,et al.  Crystal structure of human protein tyrosine phosphatase 1B. , 1994, Science.

[27]  J. Pflugrath,et al.  The finer things in X-ray diffraction data collection. , 1999, Acta crystallographica. Section D, Biological crystallography.

[28]  D. Barford,et al.  Visualization of the Cysteinyl-phosphate Intermediate of a Protein-tyrosine Phosphatase by X-ray Crystallography* , 1998, The Journal of Biological Chemistry.

[29]  J. den Hertog,et al.  Regulation of receptor protein-tyrosine phosphatase alpha by oxidative stress. , 2002, The EMBO journal.

[30]  J. Dixon,et al.  Protein tyrosine phosphatases: mechanisms of catalysis and regulation. , 1998, Current opinion in chemical biology.

[31]  B. Goldstein,et al.  Protein-tyrosine phosphatase 1B (PTP1B): a novel therapeutic target for type 2 diabetes mellitus, obesity and related states of insulin resistance. , 2001, Current drug targets. Immune, endocrine and metabolic disorders.

[32]  E. Tekle,et al.  Epidermal Growth Factor (EGF)-induced Generation of Hydrogen Peroxide , 1997, The Journal of Biological Chemistry.

[33]  D. Barford,et al.  Purification and crystallization of the catalytic domain of human protein tyrosine phosphatase 1B expressed in Escherichia coli. , 1994, Journal of molecular biology.

[34]  G. Ramponi,et al.  The Inactivation Mechanism of Low Molecular Weight Phosphotyrosine-protein Phosphatase by H2O2 * , 1998, The Journal of Biological Chemistry.

[35]  D. Barford,et al.  Structural basis for phosphotyrosine peptide recognition by protein tyrosine phosphatase 1B. , 1995, Science.

[36]  B. Neel,et al.  Protein tyrosine phosphatases in signal transduction. , 1997, Current opinion in cell biology.