| Formulation | 50%glycerol/water(v/v) |
| Storage | -20°C |
| Purity | >95%bySDS-PAGE |
| ActivityDetermination | Clottingassay |
| ShelfLife(properlystored) | 12months |
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DomainStructureofHumanFactorVIIThedomainstructureofhumanfactorVIIisrepresented,where:GLA=regioncontainingγ-carboxyglutamicacidresidues,EGF=regioncontainingsequenceshomologoustohumanepidermalgrowthfactor,CATALYTICDOMAIN=regioncontainingtheserineproteasecatalytictriad.ThesiteatwhichfactorXacleavesfactorVIItoformfactorVIIaisindicatedwithanarrow.
SampleGelInformation:
| Gel | Novex4-12%Bis-Tris |
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| Load | HumanFactorVII,1µgperlane |
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| Buffer | MOPS |
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| Standard | SeeBluePlus2;Myosin(191kDa),PhosphorylaseB(97kDa),BSA(64kDa),GlutamicDehydrogenase(51kDa),AlcoholDehydrogenase(39kDa),CarbonicAnhydrase(28kDa),MyoglobinRed(19kDa),Lysozyme(14kDa) |
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Overview:
HumanfactorVIIisasinglechain,vitaminK-dependent,plasmaglycoproteinwhichissynthesizedintheliver(1-3).Priortosecretionintotheblood,posttranslationalmodificationbyavitaminK-dependentcarboxylaseproducesten-carboxyglutamicacid(gla)residueslocatedintheNH2-terminalportionofthemolecule,whichfacilitatecellmembranebinding.FactorVIIisproteolyticallyactivatedtotheserineprotease,factorVIIa,duringcoagulation.FactorVIIcanbeactivatedbythrombin,factorIXa,factorXaorfactorXIIa.TheactivationresultsincleavageofthesinglechainmoleculeontheCOOH-terminalsideofarginine-152,toproduceanNH2-terminalderivedlightchain(Mr=20,000)andaCOOH-terminalderivedheavychain(Mr=30,000)whichremaincovalentlyassociatedbyasingledisulfidebond.Thelightchainregioncontainsthegladomain,aswellastwogrowthfactordomainswhicharehomologoustohumanepidermalgrowthfactor(EGF).Asingleβ-hydroxyasparticacididentifiedinfactorVIIisalsolocatedinthelightchainregion.TheheavychainregionoffactorVIIacontainsthecatalyticdomain.FactorVIIaandthecofactor,tissuefactor,maycombineonnegativelychargedcellsurfacesinacalciumdependentmannertoformtheextrinsicfactorXaseenzymecomplex.ThisenzymecomplexcatalyzestheconversionofbothfactorIXtofactorIXaandfactorXtofactorXa.TheCDNAforfactorVIIhasbeenisolatedandthenucleotidesequencedetermined(4).FactorVIIsharesextensivesequencehomologywithotherserineproteasesincludingfactorIX,factorXandproteinC.
HumanfactorVIIispurifiedusingacombinationofconventionaltechniques(2)andimmunoaffinitychromatography(5).Thepurifiedproteinissuppliedin50%(vol/vol)glycerol/H2Oandshouldbestoredat-20oC.PurityisdeterminedbySDS-PAGEanalysisandactivityismeasuredinafactorVIIclottingassay.
Properties:
| Localization | Plasma |
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| Plasmaconcentration | 0.5µg/ml(2)-(baseduponactivitymeasurements) |
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| Modeofaction | Zymogen;precursortotheserineproteasefactorVIIa |
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| Molecularweight | 50,000(2) |
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| Extinctioncoefficient | |
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| Isoelectricpoint | 4.8-5.1(6)-(baseduponanalysisofbovinefactorVII) |
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| Structure | singlechain,NH2-terminalgla-domain,twoEGFdomains |
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| Percentcarbohydrate | 13%(7)-(baseduponanalysisofbovinefactorVII) |
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| Post-translationalmodifications | oneβ-hydroxyaspartate(8),tenglaresidues(9) |
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