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Haematologic Technologies/Human Protein S/HCPS-0090/100 µg

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Formulation50%glycerol/water(v/v)Storage-20°CPurity>95%bySDS-PAGEActivityDeterminationN/AShelfLife(properlystored)12monthsDomainStructureofProteinSThedomainstructureofproteinSisrepresented,where:GLA=regioncontainingγ-carboxyglutamicacidresidues,EGF=regioncontainingsequenceshomologoustohumanepidermalgrowthfactor,TSR=thrombinsensitiveregion,?=regionofunknownfunctionwhichreplacesthecatalytictriadfoundinvitaminK-dependentserineproteases.SampleGelInformation:LoadHumanProteinS,1µgperlaneBufferMOPSStandardSeeBluePlus2;Myosin(191kDa),PhosphorylaseB(97kDa),BSA(64kDa),GlutamicDehydrogenase(51kDa),AlcoholDehydrogenase(39kDa),CarbonicAnhydrase(28kDa),MyoglobinRed(19kDa),Lysozyme(14kDa)Overview:ProteinSisasinglechainvitaminK-dependentproteinwhichisthoughttofunctioninboththecoagulationandcomplementcascades(1,2).Approximately60%ofproteinScirculatinginplasmaiscomplexedtoC4bbindingprotein(C4BP).Ithasbeensuggestedthatg-carboxyglutamicacid(gla)dependentbindingofproteinStonegativelychargedphospholipidsmayfunctiontoconcentrateC4BPatcellsurfacesfollowinginjury.Inthecoagulationsystem,proteinSfunctionsasananticoagulantcofactorprotein.ActivatedproteinC(APC)formsa1:1stoichiometriccomplexwithproteinSinthepresenceofCa2+andphospholipidvesicles(Kd=6x10-9M)(3).InthepresenceofproteinS,amoderateincrease(3-10fold)intherateoffactorVaandfactorVIIIainactivationbyAPCisobservedinplasmaandonthesurfaceofunstimulatedplatelets.ProteinSboundtoC4BPdoesnotpossessAPCcofactoractivity.Recently,anadditionalbindingproteinwhichenhancestheactivityofproteinShasbeendescribed(4).ProteolyticinactivationofproteinSbythrombinhasbeenproposedasaregulatorymechanisminthissystem.AsinglecleavagebythrombinabolishesproteinScofactoractivitybyremovinganNH2-terminalpeptide(Mr=8000)whichcontainsthegladomain.ThedomainstructureofproteinSissimilartothatoftheothervitaminK-dependantcoagulationfactorswiththeexceptionthatproteinSdoesnotpossessthecatalytictriad.ProteinSisasinglechainproteincontaining10glaresiduesintheNH2-terminaldomainand4epidermalgrowthfactor(EGF)domains.HumanproteinSisisolatedfromfreshfrozenplasmabyacombinationofconventionalmethods(9)andimmunoaffinitychromatographyasdescribedbyJennyetal.(5).PurifiedproteinSissuppliedin50%(vol/vol)glycerol/H2Oandshouldbestoredat-20°C.PurityisdeterminedbySDS-PAGEanalysis.Properties:LocalizationPlasma,freeandcomplexedtoC4BPPlasmaconcentration10µg/ml(free)(6)ModeofactionCofactorforactivatedproteinCMolecularweight69,000(7)ExtinctioncoefficientE1%1cm,280nm=9.5(7)Isoelectricpoint5.0-5.5(7)Structuresinglechain,NH2-terminalgladomain,fourEGFdomainsPercentcarbohydrate7%(7)Post-translationalmodificationsoneβ-hydroxyaspartate(8)tenglaresidues(7),threeβ-hydroxyasparagine(8)

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