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1、DifferentialscanningmicrocalorimetryAlanCooperChemistryDept.,GlasgowUniversity,GlasgowG128QQAdaptedfromref.21:A.Cooper,M.A.Nutley,A.Wadood,DifferentialscanningmicrocalorimetryinS.E.HardingandB.Z.Chowdhry(Eds.),Protein-LigandInteractions:hydrodynamicsandcalorimetry.OxfordUniversityPress,OxfordNe
2、wYork,(2000)p287-318.1.IntroductionDifferentialscanningcalorimetry(DSC)isanexperimentaltechniquetomeasuretheheatenergyuptakethattakesplaceinasampleduringcontrolledincrease(ordecrease)intemperature.Atthesimplestlevelitmaybeusedtodeterminethermaltransition(“melting”)temperaturesforsamplesinsoluti
3、on,solid,ormixedphases(e.g.suspensions).Butwithmoresensitiveapparatusandmorecarefulexperimentationitmaybeusedtodetermineabsolutethermodynamicdataforthermally-inducedtransitionsofvariouskinds.Formerlythiswasmoretherealmofthededicatedspecialist,butnowwiththereadyavailabilityofsensitive,stable,use
4、r-friendlyDSCinstruments,microcalorimetryhasbecomepartofthestandardrepertoireofmethodsavailabletothebiophysicalchemistforthestudyofmacromolecularconformationandinteractionsinsolutionatreasonableconcentrations.And,totheextentthatthermaltransitionsmightbeaffectedbyligandbinding,DSCcanprovideusefu
5、linformationaboutprotein-ligandbinding.Theadvantagesofcalorimetrictechniquesarisebecausetheyarebasedondirectmeasurementsofintrinsicthermalpropertiesofthesamples,andareusuallynon-invasiveandrequirenochemicalmodificationsorextrinsicprobes.Furthermore,withcarefulanalysisandinterpretation,calorimet
6、ricexperimentscandirectlyprovidefundamentalthermodynamicinformationabouttheprocessesinvolved.ThisdocumentconcentratesonthebasictheoryandpracticalapplicationsofDSCinthefieldofproteinstabilityandligandinteractions,withpracticalexamplesofitsuseanddetailsofdataanalysisandpitfalls.Itshouldbesaidfrom
7、theoutset,however,thatDSCisatbestonlyaratherindirectwayofstudyingprotein-ligandinteractions,andinmostcasesotherandmoredirectmethods(includingisothermaltitrationcalorimetry,ITC)mightbebettersuitedtotheproblem.However,thetechniqueha