HD-PTP Is a Catalytically Inactive Tyrosine Phosphatase Due to a Conserved Divergence in Its Phosphatase Domain 英文参考文献.docVIP

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HD-PTP Is a Catalytically Inactive Tyrosine Phosphatase Due to a Conserved Divergence in Its Phosphatase Domain 英文参考文献.doc

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HD-PTP Is a Catalytically Inactive Tyrosine Phosphatase Due to a Conserved Divergence in Its Phosphatase Domain 英文参考文献

HD-PTPIsaCatalyticallyInactiveTyrosinePhosphatase DuetoaConservedDivergenceinItsPhosphatase Domain Marie-ClaudeGingras1,YuLingZhang1,DmitriKharitidi1,AlastairJ.Barr2,StefanKnapp2,MichelL. Tremblay1,ArnimPause1* 1GoodmanCancer Centre and Department of Biochemistry, McGill University, Montre′al,Que′bec, Canada, 2StructuralGenomics Consortium, Nuffield Department of Medicine,UniversityofOxford,Oxford,UnitedKingdom Abstract Background: The HD-PTP protein has been described as a tumor suppressor candidate and based on its amino acid sequence, categorized as a classical non-transmembrane protein tyrosine phosphatase (PTP). To date, no HD-PTP phosphorylatedsubstratehasbeenidentifiedandcontroversialresultsconcerningitscatalyticactivityhavebeenrecently reported. MethodologyandResults:HerewereportarigorousenzymaticanalysisdemonstratingthattheHD-PTPproteindoesnot harbor tyrosine phosphatase or lipid phosphatase activity using the highly sensitive DiFMUP substrate and a panel of different phosphatidylinositol phosphates. We found that HD-PTP tyrosine phosphatase inactivity is caused by an evolutionaryconservedaminoaciddivergenceofakeyresiduelocatedintheHD-PTPphosphatasedomainsinceitsback mutationissufficienttorestoretheHD-PTPtyrosinephosphataseactivity.Moreover,inagreementwithatumorsuppressor activity,HD-PTPexpressionleadstocolonygrowthreductioninhumancancercelllines,independentlyofitscatalyticPTP activitystatus. Conclusion:Insummary,wedemonstratethatHD-PTPisacatalyticallyinactiveproteintyrosinephosphatase.Assuch,we identifyoneresidueinvolvedinitsinactivationandshowthatitscolonygrowthreductionactivityisindependentofitsPTP activitystatusinhumancancercelllines. Citation: Gingras M-C, Zhang YL, Kharitidi D, Barr AJ, Knapp S, et al. (2009) HD-PTP Is a Catalytically Inactive Tyrosine Phosphatase Due to a Conserved DivergenceinItsPhosphataseDomain.PLoSONE4(4):e5105.doi:10.1371/journal.pone.0005105 Editor:Karl-WilhelmKoch,UniversityofOldenburg,Germany ReceivedJanuary19,2009;Accepted