EFFECTS OF HYDROLASE INHIBITORS ON FERTILIZATION OF SEA-URCHINS .1. PROTEASE INHIBITORS

To search the spermatozoa of sea urchins for their lysins, the eggs were inseminated in the presence of various protease inhibitors. Among them, two chymotrypsin‐specific inhibitors, chymostatin and N‐tosyl‐L‐phenylalanyl‐chloro‐methane, as well as p‐nitrophenyl p′‐guanidinobenzoate, inhibit fertilization of the sea urchins, Hemicentrotus pulcherrimus and Strongylocentrotus intermedius. A chymotrypsin‐like protease is presumed to be a lysin of the sea urchins, since the inhibition of fertilization by chymostatin is remarkably diminished if the eggs are pretreated with trypsin or chymotrypsin to break the vitelline coat before insemination, and since N‐tosyl‐L‐phenylalanyl‐chloromethane, and p‐nitrophenyl p′‐guanidinobenzoate, as well as chymostatin, inhibit the fertilization. In all the sea urchins so far studied, elevation of fertilization envelopes is inhibited by leupeptin, antipain, soybean trypsin inhibitor, and p‐nitrophenyl p′‐guanidinobenzoate, all of which are potent trypsin inhibitors. Synthetic inhibitors have cytotoxic side effects on the eggs, but the microbial and plant inhibitors have no such effects. Copyright © 1979 Alan R. Liss, Inc.