SmCL3, a Gastrodermal Cysteine Protease of the Human Blood Fluke Schistosoma mansoni 英文参考文献.docVIP

SmCL3, a Gastrodermal Cysteine Protease of the Human Blood Fluke Schistosoma mansoni 英文参考文献.doc

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SmCL3, a Gastrodermal Cysteine Protease of the Human Blood Fluke Schistosoma mansoni 英文参考文献

SmCL3,aGastrodermalCysteineProteaseoftheHuman BloodFlukeSchistosomamansoni JanDvorˇa′k1*,SusanT.Mashiyama1,2,MohammedSajid1,3,SimonBraschi1¤a,MelaineDelcroix1¤b,EricL. Schneider2,WilsonH.McKerrow1¤c,MahmoudBahgat4,ElizabethHansell1,PatriciaC.Babbitt2, CharlesS.Craik2,JamesH.McKerrow1,ConorR.Caffrey1 1SandlerCenterforBasicResearchinParasiticDiseases,CaliforniaInstituteforQuantitativeBiosciences,UniversityofCaliforniaSanFrancisco,SanFrancisco,California, United States of America, 2Departments of Biopharmaceutical Sciences and Pharmaceutical Chemistry, California Institute for Quantitative Biosciences, University of CaliforniaSanFrancisco,SanFrancisco,California,UnitedStatesofAmerica,3LeidenUniversityMedicalCentre,afd.Parasitologie,Leiden,TheNetherlands,4Therapeutical ChemistryDepartment,InfectiousDiseasesandImmunologyLaboratory,TheRoadtoNobelProject,TheNationalResearchCenter,Dokki,Cairo,Egypt Abstract Background:BloodflukesofthegenusSchistosomaareplatyhelminthparasitesthatinfect200millionpeopleworldwide. Digestion of nutrients from the host bloodstream is essential for parasite development and reproduction. A network of proteolyticenzymes(proteases)facilitateshydrolysisofhosthemoglobinandserumproteins. Methodology/PrincipalFindings:WeidentifiedanewcathepsinLtermedSmCL3usingPCRstrategiesbasedonS.mansoni EST sequence data. An ortholog is present in Schistosoma japonicum. SmCL3 was heterologously expressed as an active enzymeintheyeast,Pichiapastoris.RecombinantSmCL3hasabroadpHactivityrangeagainstpeptidylsubstratesandis inhibitedbyClanCAproteaseinhibitors.Consistentwithafunctionindegradinghostproteins,SmCL3hydrolyzesserum albuminandhemoglobin,islocalizedtotheadultgastrodermis,andisexpressedmainlyinthoselifestagesinfectingthe mammalianhost.ThepredominantformofSmCL3intheparasiteexistsasazymogen,whichisunusualforproteases.This zymogen includes an unusually long prodomain with alpha helical secondary structure motifs. The striking specificity of SmCL3 for amino acids with

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