title: Human acrosin: purification and some properties.
abstract: Human sperm with normal morphology and good viability were obtained by centrifugation using a discontinuous Percoll density gradient with an inner column. Acrosin (E.C.3.4.21.10) was rapidly purified from sperm by ion exchange adsorption and elution and was purified by affinity adsorption on a lima bean trypsin inhibitor (LBTI) Cellulofine column. The final preparation was found to be homogeneous on polyacrylamide gel electrophoresis and to have a molecular weight of about 4 x 10(4) daltons. The enzyme had an esterolytic activity of 3.5 mumol/min/A280 with N-alpha-tosyl-L-arginine methyl ester as the substrate. Human acrosin showed a broad substrate specificity for arginine and lysine derivatives and it seemed to have a somewhat different specificity from trypsin. The optimal pH of this enzyme with amidolytic activity was 9.0. Enzyme activity was stimulated by a high concentration of calcium chloride. LBTI and aprotinin strongly suppressed the amidolytic activity with the D-valyl-L-leucyl-L-arginine-p-nitroanilide (Val-Leu-Arg-pNA) as the substrate, but alpha 1-antitrypsin and soybean trypsin inhibitor were less effective.
authors.affiliation: Department of Obstetrics and Gynecology, School of Medicine, Keio University, Tokyo, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Y
authors.initials: Y
authors.lastname: Matsuda
authors.forename: S
authors.initials: S
authors.lastname: Oshio
authors.forename: S
authors.initials: S
authors.lastname: Kaneko
authors.forename: S
authors.initials: S
authors.lastname: Nozawa
authors.forename: H
authors.initials: H
authors.lastname: Mhori
authors.forename: S
authors.initials: S
authors.lastname: Akihama
authors.forename: Y
authors.initials: Y
authors.lastname: Fujimoto
mesh_terms: D000176:Acrosin / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008297:Male; D009842:Oligopeptides / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D013094:Spermatozoa / Q000737:chemistry*; D013379:Substrate Specificity
chemical_list: D009842:Oligopeptides; D011480:Protease Inhibitors; C031006:Val-Leu-Arg-p-nitroanilide; D000176:Acrosin