Biochemical studies of the hatching process in sea urchin embryos. I. Effects of protease inhibitors

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Abstract
Effects of 12 protease inhibitors both on the hatching process and on the activity of the hatching enzyme of the sea urchins, Hemicentrotus pulcherrimus and Strongylocentrotus intermedius, were studied. Synthetic inhibitors, N‐tosyl‐L‐phenylalanyl‐chloromethane (often called tosylphenylalanine chloromethylketone; TPCK), N‐α‐tosyl‐L‐lysyl‐chloromethane (often called tosyllysine chloromethylketone; TLCK), p‐nitrophenyl p'‐guanidinobenzoate (NPGB), diphenylcarbamyl chloride (DPCC) and phenylmethane sulfonyl fluoride (PMSF), caused cytolysis of embryos, whereas the microbial inhibitors, chymostatin, leupeptin, antipain, pepstatin, elastatinal and phosphoramidon, and soybean trypsin inhibitor did not. Among them, the inhibitors specific against chymotrypsmsy chymostatin and TPCK, delayed the hatching process and inhibited the hatching enzyme. NPGB also inhibited the enzyme but its effect on the hatching process was uncertain because of its general toxicity to embryos. Esterolytic hydrolysis of N‐acetyl‐L‐tyrosine ethyl ester (ATEE) by a crude preparation of the enzyme was found.The evidence presented suggests that the hatching enzyme of sea urchins is similar to chyrmotrypsin.