title: Involvement of sperm proteases in the binding of sperm to the vitelline envelope in Xenopus laevis.
abstract: Sperm binding to the vitelline envelope in dejellied Xenopus laevis eggs was effectively inhibited by inhibitors for trypsin (soybean trypsin inhibitor and p-toluenesulfonyl-L-lysine chloroethyl ketone) and aminopeptidase B (o-phenanthroline, bestatin, and arphamenine B). Likewise, synthetic 4-methylcoumaryl-7-amide (MCA) substrates (t-butoxycarbonyl-GlyArgArg-MCA, benzyloxycarbonyl-ArgArg-MCA, and Arg-MCA) inhibited binding. Consistently, when jellied eggs were inseminated in the presence of these substrates or inhibitors for proteases, fertilization was effectively blocked. The medium in which live sperm or the sperm membrane fraction were suspended exhibited hydrolyzing activities against the synthetic substrates mentioned above, and these activities were effectively inhibited by the protease inhibitors. Ultracentrifugal fractionation of the sperm suspension following induction of the acrosome reaction by a calcium ionophore, A23187, indicated that a considerable amount of the total tryptic and aminopeptidase B activity was released into the medium. On this occasion, part of the tryptic and aminopeptidase B activity was definitely estimated to be discharged in association with a vesiculated membrane, supporting the notion that the proteases involved in binding to the vitelline envelope are present on the sperm plasma membrane.
authors.affiliation: Department of Medical Biology, Tokyo Metropolitan Institute of Medical Science, Honkomagome, Bunkyo-ku, Tokyo 113-8613, Japan. kubo@rinshoken.or.jp
authors.forename: Hideo
authors.initials: H
authors.lastname: Kubo
authors.forename: Masaharu
authors.initials: M
authors.lastname: Kotani
authors.forename: Yukio
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: Tadahiko
authors.initials: T
authors.lastname: Hazato
mesh_terms: D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D003374:Coumarins / Q000494:pharmacology; D005260:Female; D005306:Fertilization / Q000187:drug effects / Q000502:physiology; D007476:Ionophores / Q000494:pharmacology; D008297:Male; D011480:Protease Inhibitors / Q000494:pharmacology; D011485:Protein Binding; D013094:Spermatozoa / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D014817:Vitelline Membrane / Q000502:physiology*; D014982:Xenopus laevis / Q000502:physiology*
chemical_list: C502573:4-methyl-coumaryl-7-amide; D003374:Coumarins; D007476:Ionophores; D011480:Protease Inhibitors; D000001:Calcimycin