Protein Linewidth and Solvent Dynamics in Frozen Solution NMR 英文参考文献.docVIP

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Protein Linewidth and Solvent Dynamics in Frozen Solution NMR 英文参考文献.doc

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Protein Linewidth and Solvent Dynamics in Frozen Solution NMR 英文参考文献

ProteinLinewidthandSolventDynamicsinFrozen SolutionNMR AnsgarB.Siemer*¤a,Kuo-YingHuang¤b,AnnE.McDermott* DepartmentofChemistry,ColumbiaUniversity,NewYork,NewYork,UnitedStatesofAmerica Abstract Solid-stateNMRofproteinsinfrozenaqueoussolutionisapotentiallypowerfultechniqueinstructuralbiology,especiallyif itiscombined withdynamic nuclear polarization signalenhancementstrategies.Oneconcern regardingNMRstudies of frozensolutionproteinsamplesatlowtemperaturesisthattheymayhavepoorlinewidths,thuspreventinghigh-resolution studies.Tolearnmoreabouthowthesolventshellcompositionandtemperatureaffectstheproteinlinewidth,werecorded 1H, 2H,and 13Cspectraofubiquitininfrozenwaterandfrozenglycerol-watersolutionsatdifferenttemperatures.Wefound that the 13C protein linewidths generally increase with decreasing temperature. This line broadening was found to be inhomogeneous andindependent of proton decoupling. In pure water, we observe an abrupt line broadening with the freezingofthebulksolvent,followedbycontinuouslinebroadeningatlowertemperatures.Infrozenglycerol-water,wedid not observe an abrupt line broadening and the NMR lines were generally narrower than for pure water at the same temperature. 1Hand 2Hmeasurementscharacterizingthedynamicsofwaterthatisinexchangewiththeproteinshowed thatthe 13Clinebroadeningisrelativelyindependentfromthearrestofisotropicwatermotions. Citation:SiemerAB,HuangK-Y,McDermottAE(2012)ProteinLinewidthandSolventDynamicsinFrozenSolutionNMR.PLoSONE7(10):e47242.doi:10.1371/ journal.pone.0047242 Editor:AnnalisaPastore,NationalInstituteforMedicalResearch,MedicalResearchCouncil,UnitedKingdom ReceivedMay10,2012;AcceptedSeptember11,2012;PublishedOctober15,2012 Copyright: ? 2012 Siemer et al. This is an open-access article distributed under the terms of the creative commons attribution license, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:ThisworkwassupportedbyNationalScienceFoundation(