Dynamic Subunit Exchange and the Regulation of Microtubule Assembly by the Stress Response Protein Human αB Crystallin 英文参考文献.docVIP

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Dynamic Subunit Exchange and the Regulation of Microtubule Assembly by the Stress Response Protein Human αB Crystallin 英文参考文献.doc

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Dynamic Subunit Exchange and the Regulation of Microtubule Assembly by the Stress Response Protein Human αB Crystallin 英文参考文献

DynamicSubunitExchangeandtheRegulationof MicrotubuleAssemblybytheStressResponseProtein Humana BCrystallin ScottA.Houck3,JohnI.Clark1,2* 1Department of Biological Structure, University of Washington, Seattle, Washington, United States of America, 2Department of Ophthalmology, University of Washington,Seattle,Washington,UnitedStatesofAmerica,3MedicineAdministration,UniversityofNorthCarolina,ChapelHill,NorthCarolina,UnitedStatesofAmerica Abstract Background:Thesmallheatshockprotein(sHSP),humanaBcrystallin,formslarge,polydispersecomplexesthatmodulate thetubulin-microtubuleequilibriumusingadynamicmechanismthatispoorlyunderstood.TheinteractivesequencesinaB crystallinfortubulinaresurfaceexposed,andcorrespondtointeractivesitesfortheformationofaBcrystallincomplexes. Methodology/Principal Findings: There is sequence homology between tubulin and the interactive domains in the b8- strand ofthecore domainand theC-terminalextensionof aB crystallin. This studyinvestigated thehypothesis that the formationoftubulinandaBcrystallinquaternarystructureswasregulatedthroughcommoninteractivedomainsthatalter thedynamicsoftheirassembly.Sizeexclusionchromatography(SEC),SDS,microtubuleassemblyassays,aggregation assays,multiplesequencealignment,andmolecularmodelingcharacterizedthedynamicresponseoftubulinassemblyto increasingconcentrationsofaBcrystallin.LowmolarratiosofaBcrystallin:tubulinwerefavorableformicrotubuleassembly andhighmolarratiosofaBcrystallin:tubulinwereunfavorableformicrotubuleassembly.Interactionsbetweena Bcrystallin andunassembledtubulinwereobservedusingSECandSDS. Conclusions/Significance:SubunitsofaBcrystallinthatexchangedynamicallywiththeaBcrystallincomplexcaninteract withtubulinsubunitstoregulatetheequilibriumbetweentubulinandmicrotubules. Citation: Houck SA, Clark JI (2010) Dynamic Subunit Exchange and the Regulation of Microtubule Assembly by the Stress Response Protein Human aB Crystallin.PLoSONE5(7):e11795.doi:10.1371/journal.pone.0011795 Editor:StefanWo¨lfl,Universit