MMB 150 Complement Methods and Protocols

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1、MethodsinMolecularBiologyMethodsinMolecularBiologyTMVOLUME150ComplementComplementMethodsandMethodsandProtocolsProtocolsEditedbyEditedbyB.PaulMorganB.PaulMorganClassicalPathwayTerminalPathwayAlternativePathwayHUMANAPRESSHUMANAPRESSTheComplementSystem11TheComplementSystem:AnOverviewB.PaulMorgan1.Intro

2、ductionThecomplement(C)systemconsistsofagroupof12solubleplasmapro-teinsthatinteractwithoneanotherintwodistinctenzymaticactivationcas-cades(theclassicalandalternativepathways)andinthenonenzymaticassemblyofacytolyticcomplex(themembraneattackpathway)(Fig.1;Table1).Athirdactivationpathway,termedthelecti

3、npathway,hasrecentlybeendescribed(1,2).Controloftheseenzymaticcascades,essentialtopreventrapidconsumptionofCinvivo,isprovidedby10ormoreplasmaandmem-brane-boundinhibitoryproteinsactingatmultiplestagesofthesystem.Cplaysacentralroleininnateimmunedefense,whichprovidesasystemfortherapiddestructionofawide

4、rangeofinvadingmicroorganisms.ThepurposeofthisvolumeistoprovideabalancedaccountofthemethodswhichhavebeenappliedtothestudyoftheCsysteminclinicalandresearchlaboratories.Intheearlychapters,methodsfortheisolationoftheindividualCcomponents,regulatorsandreceptorswillbedescribedandassaysforthemea-suremento

5、fCactivityinthevariouspathwayswillbedetailed.LaterchapterswilldescribeaspectsofCmethodologyofrelevancetotheclinicallabora-tory—protocolsforscreeningforCdeficiencyanddeficiencyofC1inhibitor,methodsformeasurementofCactivationproductsinbiologicalfluidsandtis-suesandmethodsforallotypingCcomponents.Other

6、chapterswilldescribeadvancedtechnologieswhicharenowhavingenormousinfluenceonCresearch—structuralanalysisofthecomponentsandregulatorsandgenetargetingtogen-erateanimalsdeficientinindividualcomponentsandregulators.Inordertoappreciatethemethodologiestobedescribed,itisessentialfirsttounderstandthebasicso

7、fthesystem.From:MethodsinMolecularBiology,vol.150:ComplementMethodsandProtocolsEditedby:B.P.Morgan©HumanaPressInc.,Totowa,NJ12MorganFig.1.Thecomplementsystemanditscontrol.TheconstituentpathwaysoftheCs