资源描述：
《studies on the parameters controlling the stability of the tet peptidase》由会员上传分享，免费在线阅读，更多相关内容在工程资料-天天文库。
1、BiochimicaetBiophysicaActa1814(2011)1289–1294ContentslistsavailableatScienceDirectBiochimicaetBiophysicaActajournalhomepage:www.elsevier.com/locate/bbapapStudiesontheparameterscontrollingthestabilityoftheTETpeptidasesuperstructurefromPyrococcushorikosh
2、iirevealedacrucialroleofpHandcatalyticmetalsintheoligomerizationprocessabbb,⁎EvaRosenbaum,MylèneFerruit,M.AsunciónDurá,BrunoFranzettiaBM16-CRG,ConsorciLaboratorideLlumdeSincrotro(LLS),c/oEuropeanSynchrotronRadiationFacility,6rueJulesHorowitz,38043Greno
3、ble,FrancebInstitutdeBiologieStructuraleJ.-P.Ebel,UMR5075CNRS-GEA-UJF,41rueJulesHorowitz,38027Grenoble,FrancearticleinfoabstractArticlehistory:TheTETproteasesfromPyrococcushorikoshiiaremetallopeptidasesthatformlargedodecamericparticlesReceived3August20
4、10withhighthermalstability.Theinfluenceofvariousphysico-chemicalparametersonPhTET3quaternaryReceivedinrevisedform3November2010structurewasinvestigated.AnalyticalultracentrifugationandbiochemicalanalysesshowedthatthePhTET3Accepted24November2010quaternary
5、structureandenzymaticactivityaremaintainedinhighsaltandthatthecomplexisstableunderAvailableonline2December2010extremeacidicconditions.UnderbasicpHconditionsthecomplexdisassembledintoalowmolecularweightspeciesthatwasidentifiedasfoldeddimer.Metalanalysess
6、howedthatthepurifiedenzymeonlycontainsKeywords:Aminopeptidasetwoequivalentofzincpermonomer,correspondingtothemetalionsresponsibleforcatalyticactivity.WhenIntracellularproteolysisthesemetalswereremovedbyEDTAtreatment,thecomplexdissociatedintothesamedimer
7、icspeciesasLargemolecularcomplexesthoseobservedathighpH.DodecamericTETparticleswereobtainedfromthemetalfreedimerswhen2mMQuaternarystructureassemblingofdivalentionswereaddedtotheproteinsamples.MostofthedimersremainedassembledathighMetalbindingtemperatur
8、e.Thus,wehaveshownthatdimersarethebuildingunitsintheTEToligomerizationpathwayandHyperthermophilesthattheactivesitemetalsareessentialinthisprocess.©2010ElsevierB.V.Allrightsreserved.1.IntroductionM42accordingtotheMEROPSclassification[6].X
