Strukturbiochemie中子散射

PDDr.RegineWillumeitGKSSForschungszentrumGeesthachtMax-Planck-Str.121502GeesthachtTel.:04152871291Mobil:01704316430Email:willumeit@gkss.deStrukturbiochemieIDonnerstag,17:00–19:00Uhr10.05.200724.05.200707.06.200721.06.200705.07.2007-ExkursionzurGKSSTermineNeutronen-undRöntgenstreuung(gelösteSysteme)-3hFluoreszenzspektroskopie(verschiedeneVarianten)-2hSurfacePlasmonResonance-1hInfrarot-undCD-Spektroskopie-2hMassenspektroskopie-1hZetaPotential-1hThemenderVorlesung:AllgemeineVorbemerkungen:Strukturbiochemie-wasistdas?ErforschtdieStrukturvonProteinen,Nukleinsäuren,Lipiden,Zucker…..Komplexen(Ribosomen,Membranen,Zellen)…ErforschtdieFunktionundDynamikvonMolekülenUntersuchtdieWechselwirkungvonMolekülen(Protein)strukturanalyseDiewichtigstenDatenbankenhttp://www.rcsb.org/pdb/home/home.doEMNMRhttp://www.ncbi.nlm.nih.gov/BLAST/http://www.ncbi.nlm.nih.gov/entrezhttp://ndbserver.rutgers.edu/NDB/NDBATLAS/NucleicAcidDataBasehttp://ndbserver.rutgers.edu/NDB/index.htmlURLfunktioniertgerademalnichtLiteraturGoogle/Wikipedia!A.Pingoud&C.Urbanke:ArbeitsmethodenderBiochemie,deGryuter(1997)R.Winter&F.Noll:MethodenderbiophysikalischenChemie,Teubner(1998)Internet:PubMed:www.ncbi.nih.govScienceDirect:www.sciencedirect.comGrundlagen:WechselwirkungmitMaterieKontrastvariationAufbaueinesInstrumentesDatenaufnahmeundAuswertungStrukturanalyseanBeispielenNeutronen-undRöntgenstreuung(gelösteSysteme)AccessableSizeRegimesatomicstructuremicellespolymersproteinsvirusbacterianeutrondiffractionX-raydiffractionelectrondiffractionSANSSAXSopticalmicroscopeelectronmicroscopeatomicforcemicroscopeLSSize/mScatteringMicroscopyIntroductiontoScatteringWhattypeofscatteringdoyouknow?electromagneticwavesLightscatteringX-Rayscatteringelectromagneticwaves–shortwavelengthNeutronscatteringparticles(Electronscattering)chargedparticlesHowdotheprobesinteract?OnlyElasticScattering!InelasticscatteringTransferofenergy/momentuminto/fromthesampleatomsProbesdynamics(structuralfluctuations)MainlyusedwithneutronsInteractionofRadiationwithMatterInteractionofRadiationwithMatterInteractionwithelectronsLightscatteringInteractionwithelectronsX-rayscatteringScattering‘strength’isproportionaltoZInteractionwithelectronspinpossibleInteractionwithnuclei(protonsandneutrons)NeutronscatteringScattering‘strength’doesnotvarysystematicallyInteractionwithnuclearspinpossibleInteractionwithelectronsandelectronspinpossibleAtomicScatteringfactors/lengthHR.Winter,F.Noll:MethodenderbiophysikalischenChemie,Teubner(1998)X-RaysNeutronsatomicmass/gmol-1ComparisonNeutron-andX-ray-scatteringlengthsomerelevantelements[10-12cm]nX-ray1H-0.370.282H0.670.2812C0.661.6814N0.941.9616O0.582.2431P0.514.232S0.284.4856Fe0.956.72NeutronScatteringLengthofbiologicalrelevantelements[10-12cm][F.Sears(1986),H.GlättliundM.Goldmann(1987)]X-RaySpeciality:AnomalousScattering(ASAXS)NeutronsX-RaysIntensitylowhighH-sensitivityhighnoneIsotope-sensitivitystrongnoneHeavyelementslowhighSpin-sensitivitystrongaveragePenetrationdepthhighlowSamplesize/amountlargesmallMeasurementtimelongshortInteractionwithnucleielectronshellelectronshellunsystematicaZRadiationdamagenonehighToRemember:ToRemember:@H.Zabel,BochumContrastVariationWhenthemonstercame,Lola,likethepepperedmothandthearctichare,remainedmotionlessandundetected.Haroldofcourse,wasimmediatelydevoured.TheConceptofContrastVariationContrast=DifferenceofScatteringLengthDensitiesX-RayScatteringNeutronScatteringVolumeFractionD2OScatteringLengthDensityoftheSolvent[1010cm/cm3]ScatteringLengthDensityoftheSolute[1010cm-2]WaterSugarContrast=DifferenceofScatteringLengthDensitiesp(R)=rParticle(R)-rLM(R)p(R)=rParticle(R)-rLMr(R)=ScatteringLengthDensitiy=SumofScatteringLengthofallAtomsinaVolumeTheConceptofContrastVariationContrast=DifferenceofScatteringLengthDensitiesChangeofsolventcompositionChangeofsamplecomposition-IsotopicsubstitutionChangeofscatteringprocessUsually:CombinationofthemExamples:Polymer-SurfactantInteractionJuhaMertaandPerStenius:HelsinkiUniversityofTechnology,Espoo,FinlandVasilM.HaramusMaterials:SyntheticCationicPolymer:Poly(diallyldimethylammoniumchloride)–PDADMACM=5–50104,M~30104AnionicSurfactant:Sodiumhexadecanoate,NaPal-C15H31COONa,CAC~0.1mM,CMC=1.6mM,Kraftpoint~62oCMotivation:Howdochargedpolymersinteractwithchargedsurfactants?'Application':forexamplemodelforprotein-surfactantorDNA-surfactantinteractionusedinpaperindustriestopurifycelluloseortoimprovepaperqualityFuture:softelectronics-organicswitchesHowarethetwocomponentsorganized?InternalContrastVariation•C15H31COONa-NaPal(H)•C15D31COONa-NaPal(D)SolventD2O-10-50510152025303540contrastintensityNaPal(H)NaPal(D)PDADMAC[1010cm-2][1020cm-4]WhereisthesurfactantNaPalandwherethepolymerPDADMAC?Examples:Polymer-SurfactantInteractionSurfactantHDPDADMAC/NaPal/D2Odifferenceinthedistributionofsurfactantandpolymerintheaggregatesonthelengthscaleof20-30Å.Examples:Polymer-SurfactantInteractionInternalContrastVariationExamples:Polymer-SurfactantInteractionConclusion:ShellStructureofPDADMAC/NaPal:SurfactantCore&Polymere