Formulation	50%glycerol/water(v/v)
Storage	-20°C
Purity	>95%bySDS-PAGE
ActivityDetermination	N/A
ShelfLife(properlystored)	12months

DomainStructureofProteinZ
ThedomainstructureofproteinZisrepresented,whereGLA=theregioncontainingγ-carboxyglutamicacidresidues;EGF=theregioncontainingsequenceshomologoustohumanepidermalgrowthfactor;pseudocatalyticdomain=regionhomologoustothecatalyticchainofserineproteaseswhichlackstheactivesiteHisandSerresiduesnormallyconservedinthecatalytictriad.

SampleGelInformation:

Load	HumanProteinZ,1µgperlane
Buffer	MOPS
Standard	SeeBluePlus2;Myosin(191kDa),PhosphorylaseB(97kDa),BSA(64kDa),GlutamicDehydrogenase(51kDa),AlcoholDehydrogenase(39kDa),CarbonicAnhydrase(28kDa),MyoglobinRed(19kDa),Lysozyme(14kDa)

Overview:

HumanproteinZ(PZ)isasinglechain,vitaminK-dependantplasmaprotein(1,2).Analogouswiththemajorityofthecoagulationproteins,proteinZissynthesizedintheliver.Thematureproteincontains360aminoacids(4).BasedonaminoacidsequencehomologythedomainstructureissimilartothatofothervitaminK-dependantzymogenswhichinclude;factorVII,factorIX,factorX,andproteinC(3,4).TheN-terminalregioncontainsacarboxyglutamicacid(Gla)domainimportantinitsphospholipidmembranebindingABIlity(5).FollowingtheN-terminalGladomainaretwoEGFdomainsandaregionwhichconnectstoacatalytic-likedomain(3,4).TheC-terminalregionhasbeenshowntolackthe"typical"serineproteaseactivationsiteaswellastheHisandSerresiduesfromthecatalytictriad(3,4).Proteaseactivityhasnotbeendetectedineitherthefull-lengthproteinorcleavageproductsofproteinZ(2).FunctionallyproteinZhasbeenshowntobeadirectrequirementforthebindingofthrombintoendothelialphospholipids(6,7).ProteinZalsoservesasacofactorfortheinhibitionofcoagulationfactorXabyaplasmaserpincalledproteinZ-dependantproteaseinhibitor(ZPI)(8).InhibitionisdependantuponcomplexformationbetweenfactorXa-PZ-ZPIonthephospholipidsurface(8).

ThephysiologicalfunctionofproteinZisstillratherilldefined.Asisthecasewithothercoagulationproteinsandinhibitors,proteinZisconsumedduringdisseminatedintravascularcoagulation(DIC)(9).FurThermore,patientsdiagnosedwithaproteinZdeficiencypresentwithabnormalbleedingdiathesisduringandaftersurgicalevents(10).ThesefindingsprovidedirectevidenceastotheimportanceofproteinZinbloodcoagulation.

HumanproteinZispreparedfromfreshfrozenplasmasimilartotheproceduredescribedbyBrozeandMiletich(2).ThepurifiedproteinZissuppliedin50%(vol/vol)glycerol/H2Oandshouldbestoredat-20oC.PurityisassessedbySDS-PAGEanalysis.

Properties:

Localization	Plasma
Plasmaconcentration	2.7µg/ml(11)
Modeofaction	RequiredforthebindingofthrombintophospholipidsurfacesandforPZdependantinhibitionoffactorXa.
Molecularweight	62,000(2)
Extinctioncoefficient	E 1% 1cm,280nm =12.0(2)
Structure	Singlechain,(Mr=62,000),displaysstructuralsimilaritytoothervitaminK-dependantcoagulationfactorsbasedonsequencehomology,mostnotablyfactorX(3,4)

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