Innovative Research/ATAPhePheArgCMK/IINFFRCK/1.0mg

ProductDescription

ATA-Phe-Phe-Arg-CMK

ATA-FFRCK(Na-(acetylthio)acetyl-D-Phe-Phe-Arg-CH2Cl)andATA-FPRCK(Na-(acetylthio)acetyl-D-Phe-Pro-Arg-CHCl2)areactive-sitespecificlabelingreagentsforserineproteases.TheyarederivatizedfromirreversIBLepeptidechloromethylketonesandfacilitateincorporationofathioestermoietyviaalkylationofthecatalyticsiteHistidineresidue.Liberationofthefreethiolgroupisaccomplishedbygentletreatmentwithhydroxylamine(NH2OH)followingirreversibleincorporationintotheenzymecatalyticsite.Thefreethiolthenbecomesasiteforspecificmodificationswiththiol-reactiveprobessuchasiodoacetamidefluorescentprobes.Manyserineproteasesinwhichfreethiolsarelackingmaybespecificallylabeledattheactivesitebythesereagents.BothATA-FPRCKandATA-FFRCKhavebeenusedtolabelthrombinwith5-(iodoacetamido)fluorescein(5-IAF).Theprobewastheneffectivelyutilizedtofollowconformationalchangesinthecatalyticdomainofalpha-thrombinuponbindingtothefragment2domainofprothrombin.Inaddition,quantitativeequilibriumbindingstudiesandinvestigationsintothekineticsunderlyingthenon-proteolyticactivationofthezymogenplasminogenbystreptokinasewerecharacterizedwith2-((4-iodoacetamido)anilino)naphthalene-6-sulfonicacid(IAANS)labeledplasminogenbyusingtheATA-FFRCKreagent.References:1.Bock,P.(1993)MethodEnzymol.222:478-503.2.Bock,P.etal.(1996)JBiol.Chem.271:1072-1080.

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