Formulation	50%glycerol/water(v/v)
Storage	-20°C
Purity	>95%bySDS-PAGE
ActivityDetermination	< 100 ppm plasmin activity
ShelfLife(properlystored)	12months

HumanPlasminogen
Thedomainstructureofhumanplasminogenisrepresentedwhere:K1-K5=the5kringledomains,B-CHAIN=catalyticdomainofplasmin,andthearrowsindicatethesitesofproteolyticcleavagebyplasmin,elastase,andplasminogenactivators(PA"S).

SampleGelInformation:

Lane1	HumanGlu-Plasminogen(HCPG-0130)Reduced
Lane2	HumanGlu-PlasminogenCHOI(HCPG-0131)Reduced
Lane3	HumanGlu-PlasminogenCHOII(HCPG-0132)Reduced
Lane4	HumanLys-Plasminogen(HCPG-0133)Reduced Marker:SeeBlue+2_MOPS
Lane5	HumanGlu-Plasminogen(HCPG-0130)Non-Reduced
Lane6	HumanGlu-PlasminogenCHOI(HCPG-0131)Non-Red
Lane7	HumanGlu-PlasminogenCHOII(HCPG-0132)Non-Red
Lane8	HumanLys-Plasminogen(HCPG-0133)Non-Reduced
Gel	Novex4-12%Bis-Tris
Load	1µgperlane
Buffer	MOPS
Standard	SeeBluePlus2;Myosin(191kDa),PhosphorylaseB(97kDa),BSA(64kDa),GlutamicDehydrogenase(51kDa),AlcoholDehydrogenase(39kDa),CarbonicAnhydrase(28kDa),MyoglobinRed(19kDa),Lysozyme(14kDa)

Overview:

Plasminogenisasinglechainglycoproteinzymogenwhichissynthesizedintheliverandcirculatesinplasmaataconcentrationofapproximately2.4µM(1,2).Theplasminogenmoleculecontains790aminoacids,24disulfidebridges,nofreesulfhydrylsand5regionsofinternalsequencehomology,knownaskringles,betweenLys77andArg560.Thesefivetriple-looped,threedisulfidebridged,kringleregionsarehomologoustothekringledomainsint-PA,u-PAandprothrombin.Plasminogencontainsonehighaffinity(Kd=9x10-6M)andfourlowaffinity(Kd=5x10-3M)lysinebindingsites.Thehighaffinitybindingsiteresideswithinthefirstkringleregionofplasminogen.Theinteractionofplasminogenwithfibrinandα2-antiplasminismediatedbytheselysinebindingsites.Nativeglu-plasminogen(Mr=88,000)isreADIlyconvertedtoLys-77-plasminogen(Mr=83,000)byplasminhydrolysisoftheLys76-Lys77peptidebond.ElastasecatalyzedcleavageoftheVal441-Val442peptidebondofglu-plasminogenyieldsafunctionallyactivezymogentermedVal-442plasminogenormini-plasminogen.

Theconversionofplasminogentoplasminoccursbyavarietyofmechanisms,butallresultinhydrolysisoftheArg560-Val561peptidebondofplasminogen,yieldingtwochainswhichremaincovalentlyassociatedbyadisulfidebond.

Nativeglu-plasminogenispreparedfromfreshfrozenhumanplasmabyamodificationoftheprocedureofCastellino(3),utilizinggelfiltrationandaffinitychromatography.Thetwocarbohydratevariantsofglu-plasminogen(CHOIandCHOII)areisolatedbygradientelutionfromlysine-Sepharoseusingthelysineanalog,e-aminocaproicacid(3).Theplasminogenissuppliedin50%(vol/vol)glycerol/H2Oforstorageat-20oC.PurityisdeterminedbySDS-PAGEanalysis.

Properties:

Localization	Plasma
Plasmaconcentration	2.4µM(human)(4)
Modeofaction	Zymogen;precursortotheserineproteaseplasmin
Molecularweight	88,000(gluplasminogen)(5) 83,000(lys-plasminogen)(5) 38,000(val-plasminogen)(6)
Extinctioncoefficient	E 1% 1cm,280nm =17.0(5)
Isoelectricpoint	6.2(glu-plasminogen)(1) 6.7-8.3(lys-plasminogen)(1)
Structure	singlechain,24intrachaindisulfidebridges,5kringleregions.
Percentcarbohydrate	Approximately2%

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