最新Structural Analysis using NMR利用核磁共振结构分析-药学医学精品资料.ppt

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1、最新StructuralAnalysisusingNMR利用核磁共振结构分析-药学医学精品资料OverviewNMRstudieswereperformedinPeripherinpeptidesEpidermalGrowthfactorReceptorTransducerResultsConclusions&OutlookInteractionGARP2/PeripherinPeripherin/rds(retinaldegenerationslow):highlyconservedinbothrodandconephotorece

2、ptorsofallvertebrates4TMglycoprotein(39kDa)presentinphotoreceptoroutersegmentdiscsformshomodimersinrods(covalentlybonded),heterodimerswithROM-1arelocatedatthediscrimandmayplayaroleinanchoringthedisctothecytoskeletalsystemoftheoutersegmentTakenfromKarinPresentationCOSY(c

3、osydfesgpph)COrrelationSpectroscopYEachpairofcoupledspinsshowsupasacross-peakina2DCOSYspectrum.Thediagonalpeakscorrespondtothe1Dspectrum.Crosspeaksareusefulforassigningindividualaminoacid“spinsystems”KICYDALDPAKYAKWKPWLKPYTOCSY(dipsi2esgpph)TotalCorrelationSpectroscopyR

4、eliesonscalarorJcouplingsJcouplingbetweennucleithataremorethan3bondlengthsawayisveryweakNumberofprotonsthatcanbelinkedupina2DTOCSYspectrumisthereforelimitedtoallthoseprotonswithinanaminoacidKICYDALDPAKYAKWKPWLKPYROESY/NOESY(noesyesgpph)KICYDALDPAKYAKWKPWLKPYNuclearOverh

5、auserEnhancementSpectroscopyEachcrosspeakinaNOESYspectrumindicatesthatthenucleiresonatingatthe2frequenciesarewithin5Åinspace.IntensityofcrosspeaksisrelatedtointernucleardistanceHSQCHeteronuclearSingle-QuantumCoherencespectrumcontainsthesignalsoftheHNprotonsintheproteinb

6、ackboneEachsignalinaHSQCspectrumrepresentsaprotonthatisboundtoanitrogenatomuseoftheseheteronucleifacilitatesthestructuredetermination15N–HSQC(fhsqcf3gpph)and13C–HSQC(hsqcetgpsi2)HSQCSpectraKICYDALDPAKYAKWKPWLKPYALLKVKFDQKKRVKLAQGFigureA:1H,15N-HSQCSpectrumofPeptideP1B:1

7、H,13C-HSQCSpectrumofPeptideP2Per_P1&Garp_R2interactionPeptideP1(1.5mM)PeptideP1+R2(0.7mM)G18Contd…A.FigureA:P1overlappedonP1R215N-HSQCSpectrumB:15N-HSQCSpectrumofPeptideR2(Karin)B.ConclusionsSpectraobtainedshowwellresolvedresonances-teritiarystructureChemicalshiftsoftwo

8、residuesinP1haveshowntoshiftbymorethan0.05ppmin15NdimensionFutureWorkRunningthemissingexpt’stogetthecompleteda