| Formulation | 50%glycerol/water(v/v) |
| Storage | -20°C |
| Purity | >95%bySDS-PAGE |
| ActivityDetermination | < 100 ppm plasmin activity |
| ShelfLife(properlystored) | 12months |
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 |
| |||||
| Isoelectricpoint | 6.2(glu-plasminogen)(1) 6.7-8.3(lys-plasminogen)(1) | |||||
| Structure | singlechain,24intrachaindisulfidebridges,5kringleregions. | |||||
| Percentcarbohydrate | Approximately2% |
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