去垢剂在膜蛋白研究中的应用

©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(1,430070;2,430070):,,,,,,:,;,,,:ApplicationsofDetergentsinMembraneProteinsResearchXieHao1ChenYanke2SunEnjie1WangHongwei2(1DepartmentofBiologicalScienceandBiotechnology,WuhanUniversityofTechnology,Wuhan430070;2SchoolofResourcesandEnvironmentalEngineering,WuhanUniversityofTechnology,Wuhan430070)Abstract:Detergentsareamphiphilicmoleculesandabletosolubilisemembraneproteinsfromlipidmembranesandprovidehy2drophobicprotectionformembraneproteinsinhydrophilicsolutions.Resultantdetergent2solubilizedmembraneproteinscanmaintaintheirnativetrans2membranestructureinlipid2freeenvironments.Therefore,detergentsarevaluableinstructuralandfunctionalcharac2terizationsofmembraneproteins.Interactionsbetweendetergentsandmembraneproteinsareaffectedbytheamphiphilicfeaturestogeth2erwiththephysicalandthechemicalpropertiesofdetergents,suchascriticalMicelleconcentration.Theresearchonmembraneproteinsrequiresinvestigatorstomakefulluseofthesespecialcharacteristicsofdetergents.Ontheonehand,onecanusedetergentsinsteadoflipidmoleculestostabilizeandmaintainthestructureandactivityoflipid2freemembraneproteins;ontheotherhand,oneneedstocon2trolinteractionsbetweendetergentsandmembraneproteinsforstructuralinvestigationsonmembraneproteinsusingtechniquessuchasproteincrystallization.Inthisarticle,thelatestbibliographywerereviewedrelatedtoapplicationsofdetergentstoinvitroexpression,purification,andstructuralinvestigationofmembraneproteins.Keywords:DetergentsMembraneproteinsinvitroexpressionPurificationProteinstructure:2009211230:(30600004),:,,,:;E2mail:[e-mail],,,,,,(detergent),,,[1,2],,pH©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.()3[3],,,,()(aggregationnum2ber),()(criticalmicelleconcentration,CMC),,,,,(criticalmicelletemperature,CMT),()3,(Kraffttemperature),1,(ionicdetergents)(non2ion2icdetergents)(zwitterionicdetergents)[3,4](1),,,,,,,,,:,,;,,(lipopep2tidedetergents,LPDs),,alpha,,,[5-7]1[325]CMC(mM)DDM(n2Dodecyl2beta2D2malto2side)511011-0169850000TritonX2100625012-019100-15580000TritonX21145370135NP2400105-013OG(n2Octyl2beta2D2glucopyr2anoside)29220-258425000C12E85390111012366000n2Octyl2beta2D2thioglucopyr2anoside3089SDS(Sodiumn2dodecylsul2fate)2887-106218000CHAPS6156-104-146150LDAO(Lauryldimethylamineoxide)229269-73LPD21228390100181123100,,c,[8],,,1NupC,DDMLDAONupC,,DDMLDAONupC,602©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.:,;,,G250,G250,Bradford,,,;,,[9]2,,,,,[10-15],,NMR[10,16],,,[16],,,,,,,,,Klammt[10]EmrESugETehAYfiK,CDalpha,EmrESugETehAEmrE,EmrE,,[12,15,17,18],,(),,,,MscL,,[19],702©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.[20]3,,,Nagaraj[21]TritonX2100CHAPSSDS,,,TritonX2114,(twophasepartitioning),TritonX211430,TritonX2114,Donoghue[22,23],,,,,,,,,,,,,,,[16],(maltosebingprotein,MBP),DDMMBP,,Quick[24]Tyt1,Tyt1:(maltopyranoside),Tyt1;LDAOC12E8(glucopyranoside)Tyt1;TritonX2100Tyt1,,,,,;/;,[25],HIVVpu,[26];Toc75IIM2,[27,28],,ProFoldin,()20[29]4,802©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.[30],C12E8ATP,,ATP,ATP,ATP,,ATP,Voigt[31]II,10,NMR,,,,,NMR[4]NMR,[32-34],,,,,,,Kielec[35]KcsANMR,,X,,,,,,,,;,,//[36-38],,n2Octyl2beta2D2thiogluco2pyranoside,[39],,,Vink[40],,,Bio2Beads2beta2,[41-43],,,,,902©1994-2010ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.[7],Screpanti[44]NhaA,,;Shimizu[45],,,,,Li[46]2beta2,,5,,,,,,,,,,,,,,,,,,,,NMR;,,,,XNMR[35],[7],[1]EriksLR,MayorJA,KaplanRS.Astrategyforidentificationandquantificationofdetergentsfrequentlyusedinthepurificationofmembraneproteins.AnalyticalBiochemistry,2003,323(2):2342241.[2]ShiCW,ShaoW,XiongY,etal.Agaschromatographicmethodforquantificationofdetergentsfrequentlyusedinmembraneproteinstructuralstudies.AnalyticalBiochemistry,2008,383(2):3262328.[3]BhairiSM,MohanC.Detergents:aguidetothepropertiesandusesofdetergentsinbiologyandbiochemistry[Z].Calbiochem,EMDBio2sciences,SanDiego,CA,USA,2007.[4]SandersCR,SonnichsenF.SolutionNMRofmembraneproteins:practiceandchallenges.MagneticResonanceChemistry,2006,44:24240.[5]McGregorCL,ChenL,PomroyNC,etal.Lipopeptidedetergentsde2signedforthestructuralstudyofmembraneproteins.NatureBiotech2nology,2003,21:1712176.[6]HoDN,PomroyNC,Cuesta2SeijoJA,etal.Crystalstructureofaself2assemblinglipopeptidedetergentat1.20A.ProceedingsoftheNa2tionalAcademyofSciencesoftheUnitedStatesofAmerica,2008,105(35):1286126.[7]PriveGG.Detergentsforthestabilizationandcrystallizationofmem2braneproteins.Methods,2007,41(4):3882397.[8]QinL,HiserC,MulichakA,etal.Identificationofconservedlipid/detergent2bindingsitesi