嗜酸氧化亞鐵硫桿菌中鐵硫簇調(diào)控蛋白IscR的表達、純化及性質(zhì)研究.pdf

1、中南大學(xué)碩士學(xué)位論文嗜酸氧化亞鐵硫桿菌中鐵硫簇調(diào)控蛋白IscR的表達、純化及性質(zhì)研究姓名：張可申請學(xué)位級別：碩士專業(yè)：微生物學(xué)指導(dǎo)教師：柳建設(shè)曾嘉20080501碩士學(xué)位論文ABSTRACTABSTRACTThe[2Fe2S】clustercontainingIscRhasattractedmuchattentioninrecentyearsGeneticanalysesshowthatithasanessentialroleinthe

2、maturationofvariousironsulfurfieS)proteinsandfunctionsasacomponentofthecomplexmachineryresponsibleforthebiogenesisofFeSclustersThegeneofIscRfromAcidithiobacillusferrooxidansATCC23270wascloned，successfullyexpressedinEsche

3、richiacoliandpurifiedbyonestepaffinitychromatographytohomogenei夠ThespectraresultsoftherecombinantproteinconfirmedthattheironsulfurclusterwascorrectlyinsertedintotheactivesiteoftheproteinSite—directedmutagenesisresultsrev

4、ealedthatCys92，Cys99，andCysl07wereligatingwiththe【2Fe2S】clusteroftheproteinIronsulfur【FeS】clustersareoneofthemostubiquitousandfunctionallyversatileprostheticgroupsinnatureClusterscontain【2Fe2S】，[3Fe4S】，[4Fe4S】，or[8Fe7S】g

5、oreunitsDifferenttypesofbiological【FeS】clustershavedifferentfunctions，includingelectrontransfer，substratebinding／activation，iron／sulfurstorageregulationofgeneexpression，andenzymeactivity【FeS】clusterscouldbeincorporatedin

6、totheirproteinpartnersbothinvivoandinvitroThreedifferenttypesof[Fe—S】clusterbiosyntheticsystemshavebeendiscoveredAlthoughmostofthekeyplayersparticipatinginbiosynthesisof【FeS】clusterhavebeenidentified，themolecularmechanis

7、misstillunknownMoreover，howthedifferentproteinsinteractwitheachothertoaccomplish[FeS】proteinmaturationinvivoandhowtofaithfullyduplicatethatprocessinvitrostillneedtofurtherstudyThe【2Fe2S】clustercontainingIscRplaysanimport

8、antroleinthematurationofvariousironsulfur(FeS)proteins，andfunctionsasacomponentofthecomplexmachineryresponsibleforthebiogenesisofFe—SclustersWereportheretheassemblyof[2Fe2S】clusterinIscRfromAferrooxidansinvitrointheprese

9、nceofthreescaffoldproteinsofIscA，IscSandIscUThespectraandEPRresultsoftheholoIscRconfirmedthattheironsulfurclusterwascorrectlyassembledintotheproteinTheassemblyof【2Fe2S]clusterinIscRinvivomightalsofollowtheisc’’AUS”mechan