Matrixmetalloproteinases(MMP)areZn2+-andCa2+-dependentendopeptidases[1].MainsubfamiliesofMMPareCollagenases,gelatinases,stromelysinsandmembrane-typematrixmetalloproteinases[2].Threehomologouscollagenaseshavebeenidentifiedinhumantissues:interstitialcollagenase,neutrophilcollagenaseandcollagenase-3.Thethreeenzymescleavefibrillarcollagensatasinglesite,generatingfragmentsofapproximately¾and¼thesizeoftheoriginalmolecules.
Procollagenase-3consistsof452aminoacidswithacalculatedMrof51.680Da[3].DuetoN-linkedglycosylationtheapparentMrisabout60.000Da[4].Withintheproteinthefollowingdomainsandsequenceregionscanbedistinguished[3,4]:anN-terminalpropeptide,whichconferslatencytotheproenzyme,aCa2+-andZn2+-ionsbindingcatalyticdomain,ahingeregion,andaC-terminalhemopexin-likedomain.
Procollagenase-3canbeactivatedbyproteasesasstromelysin[4],gelatinaseA,membranetype1matrixmetalloproteinaseandplasmin[5]orbyincubationwithorganomercurials(e.g.APMA)[4].
Activecollagenase-3beginswiththeN-terminalsequenceYNVFPRTL[4].Collagenase-3hydrolyzestypeIIcollagen5-to6-timesfasterthantypeIandtypeIIIcollagens.TheenzymeexhibitsalsohighactivitytowardsgelatinanditdegradesSERPINSas1-antichymotrypsinandplaminogenactivatorinhibitor-2[4].Collagenase-3isinhibitedina1:1stoichiometricfashionbyTIMP-1,TIMP-2andTIMP-3.Collagenase-3isexpressedduringfetalbonedevelopment[6]. Inadulthumantissuescollagenase-3hasbeendetectedonlyinpathologicalconditions:inmalignanttumors[3],inchroniculcers[7],inarthriticcartilage[8]andsynovium[9].
Purity:Recombinantprocollagenase-3appearsasasingleproteinbandofabout60.000DainSDS-PAGE(>95%oftotalprotein).Duetoautoproteolyticactivityminorbandsofactivatedcollagenase-3maybevisIBLeintheenzymepreparation.
SpecificActivity:Thespecificactivityofcollagenase-3is≥200mU/mg.1Uistheactivitythathydrolyzes1molpeptide(7-methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Leu-(3-[2,4-dinitrophenyl]-L-2,3-diaminopropionyl)-Ala-Arg-NH2(Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg)within1minundertheassayconditionsdescribedbelow.
Inhibitors:Collagenase-3isinhibitedbytissueinhibitorsofmatrixmetalloproteinases(TIMP)andbychelatorsofdivalentcationsasEDTAoro-phenanthroline.
StABIlity&Storage:Recombinantprocollagenase-3tableuntiltheexpirydategivenonthelabelifstoredat-70°C.Theproenzymecanbekeptat-20°Cforseveralweeks.Repeatedfreezingandthawingshouldbeavoided.
Applications:Recombinantprocollagenase-3canserveasantigenstandardinimmunochemicalanalyses. Theactiveenzymemaybeusedtostudythedegradationofextracellularmatrixproteins,toscreeninhibitorsofmatrixmetalloproteinasesandtocharacterizeinhibitoractions.
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植物：抗虫、抗病、抗除草剂植物培育、改良作物品质（富含赖氨酸的玉米）
动物：提高生长速度、生产药物蛋白、器官移植、乳腺生物反应器
微生物：疫苗
基因治疗等等
各位业内前辈，我们正在考虑引进符合GMP认证标准的CHO细胞系，用于表达可做疫苗生产的重组蛋白类。目前已有符合标准信息的是Thermofisher的CHO-S悬浮培养细胞资料，希望能再多了解一些和这株细胞类似的其他公司符合GMP标准的生产株细胞做个比较。谢谢指教！
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生物合成人胰岛素注射液，比如诺和灵R笔芯。是单一品种、不是混合的胰岛素。是不含鱼精蛋白的短效胰岛素。
