生物专业英语综述作业Word格式文档下载.docx

生物专业英语综述作业Word格式文档下载.docx

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ThesiteandthemechanismofoxidationofArgininekinase

Abstract

Argininekinase（argininekinase,AK）（EC2.7.3.3）isaphosphodiesteraseguanidineskinase,composedof403aminoacids,whichcontainsfivecysteines（Cys23,Cys127,Cys139,Cys201,Cys271）,theexperimentwefoundthattheoxidationstatebandandrestorethestatewiththephenomenonofelectrophoresisAK,andthroughaseriesofbiotechnologymeans,respectively,ofthesefivesitesCysmutatedtoserine（Ser）,toidentifyoxidationsitesandtostudytheoxidationmechanism,andbyUVfluorescence,respectively,toexplorethestructureafterthemutationofthetheAKvarioussitesrelatedtoactivity.

Keywords:

Argininekinase,rite-directedmutation,oxidationsites,SDS-PAGE

Introduction

1.Argininekinase[1]（ArgininekinaseAK）（EC2.7.3.3）isakinaseofaphosphorylatedoriginalguanidinocompoundexistswidelyininvertebrates,andisanimportantkinasethathasadirectrelationshipwithintracellularenergyoperation,musclecontraction,ATP-regenerating.ItsFunctionishowtoCatalyticreversiblereactionsbythefollowingstep:

ATPphosphategroupistransferredtothearginine,therebyformingahighenergybondenergystoragemolecules-argininephosphate.

Thereactionequationisasfollows:

Mg2+ATP+ArgMg2+ADP+Argininephosphate

earlyin1998,MrZhouwithsomepeoplehaveusedthecrystalstructureofthetransitionstateAKfromhorseshoecrabsbyhorseshoetocrabtheLimuluspolyphemussinglesubunitAKdissociationcombinedtransitionstateanalogs.TheresultsshowedthattheAKwasmadeupbyasmallα-helixoftheN-terminaldomainandalargeC-terminalstructure（112,-357residues）.TheC-terminaldomainisSimilartotheC-terminaldomainofglutamatesynthase,thetheBaguantiparallelβ-sheetisaroundbysevenα-helicalpackage[2].Seenfromthisstructure,thetransitionstateanalogisnotincorporatedinthemiddleofthetwodomains,butcombinedintheC-terminal'

sInteriorregion,whichmeansthattheAKistheactivesiteintheCterminal.Inaddition,theC-terminaldomaincanbedividedintotwosmallstructure,whileatransitionstateanalogsisincorporatedatthejunctionofthetwodomains[3].AccordingtoMr.ZouChenglu'

spointofviewthattheactivesiteoftheenzymeisinarelativelyflexibleportionandisthepartsofthestructure,andthisPositionismoresusceptiblefromtheimpactofadverseexternalfactorsthanotherparts,theresultisthattheenzymeactivitylossfasterandeasierthantheenzymeofthewholestructure.

2.Theoxidationofprotein

Differenttypesofoxidativedamageinproteinisanimportantfacterofthecauseofmanyseriousdiseases（suchas:

cancer,diabetes,neurodegenerativediseases）ThechaperoneproteinoxidativedamageofER,willleadtothemisfoldingandaggregationdiseasesinprotein,suchas:

AD,Parkinson'

sdisease（Parkinson'

sdisease,PD）,amyotrophiclateralsclerosis（amyotrophicoflateralsclerosis,ALS）,andHangextensionRatonchorea（Huntington'

sdisease,HD）,andsenescence-relateddiseases,animportantcommonfeatureofthemistheabnormalproteininthebrainwithintheaccumulation.

IntheexperimentofelectrophoreticdetectionwithAKwefoundtheoxidationstatebandandreducedstateband.Revelatedbytheresearchmethodsofcreatinekinase（CK）,wemake5sitesCysofAKmutatedtoserine（Ser）,thenuseofaffinitychromatography,UV,fluorescence,suchasaran