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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.
T Kobayashi, and
Y Matsuda, and
S Oshio, and
S Kaneko, and
S Nozawa, and
H Mhori, and
S Akihama, and
Y Fujimoto
November 1973,
European journal of biochemistry,
T Kobayashi, and
Y Matsuda, and
S Oshio, and
S Kaneko, and
S Nozawa, and
H Mhori, and
S Akihama, and
Y Fujimoto
December 1980,
Hoppe-Seyler's Zeitschrift fur physiologische Chemie,
T Kobayashi, and
Y Matsuda, and
S Oshio, and
S Kaneko, and
S Nozawa, and
H Mhori, and
S Akihama, and
Y Fujimoto
January 1982,
Canadian journal of biochemistry,
T Kobayashi, and
Y Matsuda, and
S Oshio, and
S Kaneko, and
S Nozawa, and
H Mhori, and
S Akihama, and
Y Fujimoto
December 1972,
Hoppe-Seyler's Zeitschrift fur physiologische Chemie,
T Kobayashi, and
Y Matsuda, and
S Oshio, and
S Kaneko, and
S Nozawa, and
H Mhori, and
S Akihama, and
Y Fujimoto
January 1973,
Proceedings of the National Academy of Sciences of the United States of America,
T Kobayashi, and
Y Matsuda, and
S Oshio, and
S Kaneko, and
S Nozawa, and
H Mhori, and
S Akihama, and
Y Fujimoto
July 1981,
Chemical & pharmaceutical bulletin,
T Kobayashi, and
Y Matsuda, and
S Oshio, and
S Kaneko, and
S Nozawa, and
H Mhori, and
S Akihama, and
Y Fujimoto
October 1986,
Biochemistry and cell biology = Biochimie et biologie cellulaire,
T Kobayashi, and
Y Matsuda, and
S Oshio, and
S Kaneko, and
S Nozawa, and
H Mhori, and
S Akihama, and
Y Fujimoto
January 1985,
Zeitschrift fur medizinische Laboratoriumsdiagnostik,
T Kobayashi, and
Y Matsuda, and
S Oshio, and
S Kaneko, and
S Nozawa, and
H Mhori, and
S Akihama, and
Y Fujimoto
September 1976,
Biochimica et biophysica acta,
T Kobayashi, and
Y Matsuda, and
S Oshio, and
S Kaneko, and
S Nozawa, and
H Mhori, and
S Akihama, and
Y Fujimoto
July 1972,
Biochimica et biophysica acta,
