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• W1623774436 endingPage "909" @default.
• W1623774436 startingPage "903" @default.
• W1623774436 abstract "We report a one-step method for the purification to homogeneity of a cysteine proteinase of Entamoeba histolytica (histolysin) by affinity chromatography of the soluble extract of the parasite on immobilized phenylalanyl(2-phenyl)aminoacetaldehyde semicarbazone. The enzyme has an apparent Mr of 26,000 by SDS/polyacrylamide-gel electrophoresis and 29,000 by gel chromatography. Its pH optimum varies widely, from 5.5 with azocasein to approx. 7 with other protein substrates and benzyloxycarbonylphenylalanyl-L-citrullylaminomethylcourmarin++ + (Z-Phe-Cit-NHMec), and to 9.5 with benzyloxycarbonylphenylalanylarginylaminomethylcoumarin (Z-Phe-Arg-NHMec) and benzyloxycarbonylarginylarginylaminomethylcourmarin (Z-Arg-Arg-NHMec). Values of Km, kcat. and kcat/Km are 1.5 microM, 130 s-1 and 87 X 10(6) M-1.s-1 for Z-Arg-Arg-NHMec, and 32 microM, 0.4 s-1 and 0.012 x 10(6) M-1.s-1 for Z-Phe-Arg-NHMec, respectively, at pH 7.5 and 37 degrees C. The enzyme is inhibited by leupeptin and such inhibitors of cysteine proteinases as L-transepoxysuccinyl-L-leucylamido-4-(guanidino)butane, peptidyldiazomethanes, iodoacetic acid and chicken cystatin. The tentative N-terminal amino acid sequence of the enzyme closely resembles that of papain. Histolysin does not degrade type I collagen or elastin, but it is active against cartilage proteoglycan and kidney glomerular basement-membrane collagen. It also detaches cells from their substratum in vitro, and could well play a role in tissue invasion." @default.
• W1623774436 created "2016-06-24" @default.
• W1623774436 creator A5018793100 @default.
• W1623774436 creator A5032943700 @default.
• W1623774436 date "1988-03-15" @default.
• W1623774436 modified "2023-09-27" @default.
• W1623774436 title "Affinity purification and biochemical characterization of histolysin, the major cysteine proteinase of <i>Entamoeba histolytica</i>" @default.
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• W1623774436 doi "https://doi.org/10.1042/bj2500903" @default.
• W1623774436 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/1148941" @default.
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• W1623774436 hasPublicationYear "1988" @default.
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