HUMAN ACROSIN - PURIFICATION AND SOME PROPERTIES

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-mu-mol/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 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.