Effects of protease inhibitors on binding of sperm to the vitelline coat of ascidian eggs: implications for participation of a proteasome (multicatalytic proteinase complex).
Among various protease inhibitors, chymostatin (an inhibitor of sperm chymotrypsin-like protease) strongly inhibited the binding of sperm to the vitelline coat of glycerinated eggs of the ascidian Halocynthia roretzi, whereas leupeptin (an inhibitor for sperm acrosin), antipain, and soybean trypsin inhibitor had no significant inhibitory effects. Dansyl-Val-Pro-argininal (an inhibitor of the sperm trypsin-like protease, spermosin) had an inhibitory effect on the binding of sperm that was much smaller than its effects on fertilization. Since the sperm chymotrypsin-like protease that is involved in ascidian fertilization has been identified as a proteasome (multicatalytic proteinase complex), we tested the effects of several peptidyl argininals, inhibitors of the activities of proteasomes, on this binding process. The ranking of the inhibitory effects of these compounds on the binding of sperm was the same as that of their effects on the chymotrypsin-like activity of the proteasome, reported previously. The potent inhibitors of binding used in these studies had no or minimal effects on sperm motility. These results suggest that a sperm chymotrypsin-like protease (most probably the chymotrypsin-like protease in the proteasome) plays a key role in binding of sperm to the vitelline coat of the ascidian egg.
Takizawa
S
S
Department of Life Science, Tokyo Institute of Technology, Nagatsuta, Yokohama.
Sawada
H
H
Someno
T
T
Saitoh
Y
Y
Yokosawa
H
H
Hoshi
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives; D003546:Cysteine Endopeptidases / Q000502:physiology*; D005260:Female; D005306:Fertilization / Q000502:physiology; D008297:Male; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000502:physiology*; D009842:Oligopeptides / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology*; D046988:Proteasome Endopeptidase Complex; D013081:Sperm Motility; D013084:Sperm-Ovum Interactions / Q000187:drug effects / Q000502:physiology*; D014561:Urochordata; D014817:Vitelline Membrane / Q000187:drug effects / Q000502:physiology*
D009097:Multienzyme Complexes; D009842:Oligopeptides; D011480:Protease Inhibitors; D001120:Arginine; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex