Dynamics of Tropomyosin in Muscle Fibers as Monitored by Saturation Transfer EPR of Bi-Functional Probe 英文参考文献.docVIP

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Dynamics of Tropomyosin in Muscle Fibers as Monitored by Saturation Transfer EPR of Bi-Functional Probe 英文参考文献.doc

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Dynamics of Tropomyosin in Muscle Fibers as Monitored by Saturation Transfer EPR of Bi-Functional Probe 英文参考文献

DynamicsofTropomyosininMuscleFibersasMonitored bySaturationTransferEPRofBi-FunctionalProbe RoniF.Rayes1,2,Tama′sKa′lai3,Ka′lma′nHideg3,MichaelA.Geeves4,PiotrG.Fajer1,2* 1InstituteofMolecularBiophysics,FloridaStateUniversity,Tallahassee,Florida,UnitedStatesofAmerica,2NationalHighMagneticFieldLaboratory,Tallahassee,Florida, UnitedStatesofAmerica,3DepartmentofOrganicandMedicinalChemistry,UniversityofPe′cs, Pe′cs,Hungary,4SchoolofBiosciences,UniversityofKent,Canterbury, UnitedKingdom Abstract Thedynamicsoffourregionsoftropomyosinwasassessedusingsaturationtransferelectronparamagneticresonanceinthe muscle fiber. In order to fully immobilize the spin probe on the surface of tropomyosin, a bi-functional spin label was attached to i,i+4 positions via cysteine mutagenesis. The dynamics of bi-functionally labeled tropomyosin mutants decreasedbythreeordersofmagnitudewhenreconstitutedinto‘‘ghostmusclefibers’’.Theratesofmotionvariedalong the length of tropomyosin with the C-terminus position 268/272 being one order of magnitude slower then N-terminal domainorthecenterofthemolecule.Introductionoftroponindecreasesthedynamicsofallfoursitesinthemusclefiber, buttherewasnosignificanteffectuponadditionofcalciumormyosinsubfragment-1. Citation: Rayes RF, Ka′lai T, Hideg K, Geeves MA, Fajer PG (2011) Dynamics of Tropomyosin in Muscle Fibers as Monitored by Saturation Transfer EPR of Bi- FunctionalProbe.PLoSONE6(6):e21277.doi:10.1371/journal.pone.0021277 Editor:AnnalisaPastore,NationalInstituteforMedicalResearch,MedicalResearchCouncil,London,UnitedKingdom ReceivedMarch17,2011;AcceptedMay24,2011;PublishedJune20,2011 Copyright: ? 2011 Rayes 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:ThisworkwasfundedbytheHungarianResearchFundOTKAK81123toTKandKH,andtheNSFMCP0346650granttoRRandPF.Thefundershadno r