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Haematologic Technologies/Human Plasmin/HCPM-0140/500 µg

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HCPM-0140

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Formulation50%glycerol/water(v/v)Storage-20°CPurity>95%bySDS-PAGEActivityDeterminationChromogenicassayShelfLife(properlystored)12monthsHumanPlasminogenThedomainstructureofhumanplasminogenisrepresentedwhere:K1-K5=the5kringledomains,B-CHAIN=catalyticdomainofplasmin,andthearrowsindicatethesitesofproteolyticcleavagebyplasmin,elastase,andplasminogenactivators(PA"S).SampleGelInformation:GelNovex4-12%Bis-TrisLoadHumanPlasmin,1µgperlaneBufferMESStandardSeeBluePlus2;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:LocalizationPlasmaModeofactionEnzymeinvolvedinfibrinolysisMolecularweight83,000(lys-plasmin)(4)ExtinctioncoefficientE1%1cm,280nm=17.0(4)Isoelectricpoint6.7-8.3(3)StructureTwosubunits,Mr=57,000andMr=26,000,disulfidelinked,5kringledomains22disulfidebridges,NH2-terminallysine.PercentcarbohydrateApproximately2%(2)

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