Secreted Protein Acidic and Rich in Cysteine Is a Matrix Scavenger Chaperone 英文参考文献.docVIP

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Secreted Protein Acidic and Rich in Cysteine Is a Matrix Scavenger Chaperone 英文参考文献.doc

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Secreted Protein Acidic and Rich in Cysteine Is a Matrix Scavenger Chaperone 英文参考文献

SecretedProteinAcidicandRichinCysteineIsaMatrix ScavengerChaperone AlexandreChlenski1*,LisaJ.Guerrero1,HelenR.Salwen1,QiweiYang1¤a,YufengTian1¤b ,Andres MoralesLaMadrid1,SalidaMirzoeva2,PatriceG.Bouyer3,DavidXu1,MatthewWalker4¤c,SusanL.Cohn1 1DepartmentofPediatrics,UniversityofChicago,Chicago,Illinois,UnitedStatesofAmerica,2DepartmentofPathology,NorthwesternUniversity,Chicago,Illinois,United States ofAmerica, 3Department ofSurgery,University ofChicago,Chicago, Illinois, United States ofAmerica, 4Committee onCancer Biology,University ofChicago, Chicago,Illinois,UnitedStatesofAmerica Abstract SecretedProteinAcidicandRichinCysteine(SPARC)isoneofthemajornon-structuralproteinsoftheextracellularmatrix (ECM) in remodeling tissues. The functional significance of SPARC is emphasized by its origin in the first multicellular organismsanditshighdegreeofevolutionaryconservation.AlthoughSPARChasbeenshowntoactasacriticalmodulator ofECMremodelingwithprofoundeffectsontissuephysiologyandarchitecture,noplausiblemolecularmechanismofits actionhasbeenproposed.Inthepresentstudy,wedemonstratethatSPARCmediatesthedisassemblyanddegradationof ECMnetworksbyfunctioningasamatricellularchaperone.Whileithaslowaffinitytoitstargetsinsidethecellswherethe Ca 2+ concentrationsarelow,highextracellularconcentrationsofCa2+activatebindingtomultipleECMproteins,including collagens.Wedemonstratedthatinvitro,thisleadstotheinhibitionofcollagenIfibrillogenesisanddisassemblyofpre- formedcollagenIfibrilsbySPARCathighCa concentrations.Incellculture,exogenousSPARCwasinternalizedbythe 2+ fibroblastcellsinatime-andconcentration-dependentmanner.Pulse-chaseassayfurtherrevealedthatinternalizedSPARC is quickly released outside the cell, demonstrating that SPARC shuttles between the cell and ECM. Fluorescently labeled collagen I, fibronectin, vitronectin, and laminin were co-internalized with SPARC by fibroblasts, and semi-quantitative WesternblotshowedthatSPARCmediatesinternalizationofcollagenI.Usinganovel3-dimention