Functional Analyses of Complement Convertases Using C3 and C5-Depleted Sera 英文参考文献.docVIP

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Functional Analyses of Complement Convertases Using C3 and C5-Depleted Sera 英文参考文献.doc

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Functional Analyses of Complement Convertases Using C3 and C5-Depleted Sera 英文参考文献

FunctionalAnalysesofComplementConvertasesUsing C3andC5-DepletedSera MarcinOkroj,EmelieHolmquist,BenC.King,AnnaM.Blom* DepartmentofLaboratoryMedicine,LundUniversity,TheWallenbergLaboratory,Malmo¨,Sweden Abstract C3andC5convertasesarecentralstagesofthecomplementcascadesincetheyconvergethedifferentinitiationpathways, augment complement activation by an amplification loop and lead to a common terminal pathway resulting in the formation of the membrane attack complex. Several complement inhibitors attenuate convertase formation and/or acceleratedissociationofconvertasecomplexes.Functionalassaysusedtostudytheseprocessesareoftenperformedusing purified complement components, from which enzymatic complexes are reconstituted on the surface of erythrocytes or artificial matrices. This strategy enables identification of individual interactions between convertase components and putativeregulatorsbutcarriesaninherentriskofdetectingnon-physiologicalinteractionsthatwouldnotoccurinamilieu ofwholeserum.Herewedescribeanovel,alternativemethodbasedonC3orC5-depletedsera,whichsupportactivationof thecomplementcascadeuptothedesiredstagesofconvertases.Thisapproachallowsfastandsimpleassessmentofthe influenceofputativeregulatorsonconvertaseformationandstability.Asanexampleofpracticalutilityoftheassay,we performedstudiesonthioredoxin-1inordertoclarifythemechanismofitsinfluenceoncomplementconvertases. Citation: OkrojM, HolmquistE,KingBC,BlomAM(2012)Functional AnalysesofComplementConvertasesUsing C3andC5-DepletedSera.PLoSONE 7(10): e47245.doi:10.1371/journal.pone.0047245 Editor:AnnekeI.denHollander,RadboudUniversityNijmegenMedicalCentre,TheNetherlands ReceivedJune4,2012;AcceptedSeptember11,2012;PublishedOctober10,2012 Copyright: ? 2012 Okroj 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:Thisworkwassu