来自于变性链球菌的脱氧胞嘧啶核苷酸脱氨酶的表达，纯化，结晶以及初步晶体学研究.docVIP

来自于变性链球菌的脱氧胞嘧啶核苷酸脱氨酶的表达，纯化，结晶以及初步晶体学研究 生物化学与生物物理进展ProgressinBiochemistryandBhysics,2006,33(7):673--,676 ,)ln)nⅣ.pibb.ac.ca PIlaf! Expression,Purification,Crystallizationand PreliminaryX—rayStudiesofaDe0xycytidylate DeaminaseFromStreptococcusmutans霉 HOUHai—Feng,GAOZeng-Qiang,XUJian—Hua,XURuiLILi—Qin,LILan—Fen3), LLNGYu_He,SUXiao—Dong3),LI【JPeng,ⅫNDing-Chang,DONGYu-Hui (讽SynchrotronRadiationFacility,InstituteofHighEnergyPhysics,TheChineseAcademyofSciences,Be~ing100049,China; ZGraduateSchoolofTheChineseAcademyofSciences,Be堍100049China; 3~Collegeof￡Science,拍Universamp;100871,China) AbstractDeoxycytidylate(dCMP)deaminaseisallb~lzylnebelongedtodl姗cytdeamfamily.ThedCNdeaminasefrom Streptococcusma,UAl59wasclonedandexpressedin最coli.andpurifiedtohomogeneity.TheFPLCsizeexclusion chromatographyanalysisrevealsthatthes-mI/~on$dCNdeaminaseformshexamerinsolution.Theproteinwascrystallizedusing hangingdropvaponr-diffusionmethodanddiffractedtoaresolutionof3.1A.ThediffractiondatawerecollectedatBSRFbeamline 3wlA.ThecrystalsbelongtoP2I3spacegroup,withunitcellparametersa=b=c=ll3.2A,ot=卢=7=90..Assumingtherearetwo subunitsperasymmetricunit.theMatthewsCO0clentis3.6ADa-1.Thisisthefirstcrystallizationreportofthewild-type deoxycytidylatedeaminase. Keywordsdeoxycytidylatedeaminase,Streptococcusma,,allostcricregulation,crystallization Wild-typedeoxycytidylatedeaminase,a hexamericallosterice],mebindingzincion,whichis requiredforcatalyticactivity,belongstodCⅣ【Ecyt deamfamilyaccordingtoPfamsearcht.Theactivity oftheenzymeisalIostericallvregulatedbytheratio0f dC:TPtod1n渊.notonlyineukaryoticcellsbutalso inT.evenphage—infectedEscherichiacoli.withdC1]P actingasanactivatorandd1门asaninhibitor. Deoxycytidylatedeaminasecatalyzesthedeamination ofdCⅣ【EtodI.providingthenucleotidesubstrate forthymidylatesynthase,whichisimportantin DNAsynthesisandcancerchemotherapvM.dCⅣ【E deaminaseexistsinmosteukaryoticandbacterial organisms.TherearetwoclassesinthesedCⅣ【E deaminasesC3.5.4.12).MammaliandCⅣ【E deaminasecontainsasing