F- and G-Actin Concentrations in Lamellipodia of Moving Cells 英文参考文献.docVIP

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F- and G-Actin Concentrations in Lamellipodia of Moving Cells 英文参考文献.doc

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F- and G-Actin Concentrations in Lamellipodia of Moving Cells 英文参考文献

F-andG-ActinConcentrationsinLamellipodiaofMoving Cells StefanA.Koestler1,KlemensRottner2,FrankLai2,JenniferBlock2,MarleneVinzenz1,J.VictorSmall1* 1InstituteofMolecularBiotechnology, AustrianAcademyofSciences,Vienna,Austria, 2CytoskeletonDynamicsGroup,HelmholtzCentreforInfectionResearch(HZI), Braunschweig,Germany Abstract Cellsprotrudebypolymerizingmonomeric(G)intopolymeric(F)actinatthetipofthelamellipodium.Actinfilamentsare depolymerizedtowardstherearofthelamellipodiuminatreadmillingprocess,therebysupplementingaG-actinpoolfora new round of polymerization. In this scenario the concentrations of F- and G-actin are principal parameters, but have hitherto not been directly determined. By comparing fluorescence intensities of bleached and unbleached regions of lamellipodiainB16-F1mousemelanomacellsexpressingEGFP-actin,beforeandafterextractionwithTritonX-100,weshow thattheratioofF-toG-actinis3.2+/20.9.UsingelectronmicroscopytodeterminetheF-actincontent,thisratiotranslates intoF-andG-actinconcentrationsinlamellipodiaofapproximately500mMand150mM,respectively.TheexcessofG-actin, atseveralordersofmagnitudeabovethecriticalconcentrationsatfilamentendsindicatesthatthepolymerizationrateis notlimitedbydiffusionandistightlycontrolledbypolymerization/depolymerizationmodulators. Citation:KoestlerSA,RottnerK,LaiF,BlockJ,VinzenzM,etal.(2009)F-andG-ActinConcentrationsinLamellipodiaofMovingCells.PLoSONE4(3):e4810. doi:10.1371/journal.pone.0004810 Editor:NeilHotchin,UniversityofBirmingham,UnitedKingdom ReceivedNovember27,2008;AcceptedJanuary27,2009;PublishedMarch11,2009 Copyright: ? 2009 Koestler 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:HumanFrontierScienceProgramOrganisation(HFSPO);TheViennaScienceResearchandTechnologyFund(WWTF);TheCityofVienna/Zentrumfu¨r Innovation und Technologie via the