| Formulation | 50%glycerol/water(v/v) |
| Storage | -20°C |
| Purity | >95%bySDS-PAGE |
| ActivityDetermination | Chromogenicassay |
| ShelfLife(properlystored) | 12months |
Thedomainstructureofhumanplasminogenisrepresentedwhere:K1-K5=the5kringledomains,B-CHAIN=catalyticdomainofplasmin,andthearrowsindicatethesitesofproteolyticcleavagebyplasmin,elastase,andplasminogenactivators(PA"S).
SampleGelInformation:
| Gel | Novex4-12%Bis-Tris |
|---|---|
| Load | HumanPlasmin,1µgperlane |
| Buffer | MES |
| Standard | SeeBluePlus2;Myosin(191kDa),PhosphorylaseB(97kDa),BSA(64kDa),GlutamicDehydrogenase(51kDa),AlcoholDehydrogenase(39kDa),CarbonicAnhydrase(28kDa),MyoglobinRed(19kDa),Lysozyme(14kDa) |
Overview:
Plasminogen(whetherGlu-1,Lys-77orVal-442)isconvertedtotheactiveserineproteaseplasminbyhydrolysisoftheArg560-Val561peptidebondyieldinganNH2-terminalheavy(A)chainandaCOOH-terminallight(B)chainlinkedby2disulfidebonds(1-3).Thisconversioniscatalyzedbyavarietyofphysiologicalandpathologicalactivators,includingurinarytypeplasminogenactivators,tissuetypeplasminogenactivators,streptokinase,staphylokinase,kallikrein,factorsIXaandXIIa.TheCOOH-terminalderivedlightchain(Mr=26,000)containsthecatalytictriad(His42,Asp85andSer180)aswellasthestreptokinasebindingsite.TheNH2-terminalderivedheavychainrangesinmolecularweightfrom63,000to12,000dependingonthetypeofplasminogenfromwhichitoriginated.Intheabsenceofinhibitors,plasmincleavestheamino-terminalGlu1toLys76peptidefromplasmin(plasminogen)toyieldLys-plasmin,whichhasagreateraffinityforfibrinthantheGluform.TheheavychainofLys-plasminogencontains5tripleloopdisulfidebridgedregionsofinternalsequencehomologyknownaskringles.Kringles1-4containtheω-aminocarboxylicacidandfibrinbindingsites.
Plasminisaserineproteasewithbroadspecificitywhich,inadditiontocleavageoffibrin,iscapableofactivationand/ordegradationofcompoundsofthecoagulation,kiningenerationandcomplementsystems.Althoughplasmincanbeinhibitedbyanumberofplasmaproteaseinhibitorsinvitro,regulationofplasmininvivoisthoughttooccurmainlythroughitsinteractionwithα2-antiplasmin,andtoalesserextent,α2-macroglobulin.
HumanLys-plasminispreparedfromhomogeneousGlu-plasminogenusingurokinase,asdescribedbyRobbinsetal.(3).Plasminissuppliedin50%(vol/vol)glycerol/H2Oandshouldbestoredat-20°C.ActivityismeasuredbychromogenicsubstrateassayandpurityisjudgedbySDSgelelectrophoresis.
Properties:
| Localization | Plasma | |||||
|---|---|---|---|---|---|---|
| Modeofaction | Enzymeinvolvedinfibrinolysis | |||||
| Molecularweight | 83,000(lys-plasmin)(4) | |||||
| Extinctioncoefficient |
| |||||
| Isoelectricpoint | 6.7-8.3(3) | |||||
| Structure | Twosubunits,Mr=57,000andMr=26,000,disulfidelinked,5kringledomains22disulfidebridges,NH2-terminallysine. | |||||
| Percentcarbohydrate | Approximately2%(2) |
