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Properties of Intracellular Hatching enzyme in the Embryos of the Sea Urchin, Hemicentrotus pulcherrimus
Author(s) -
NAKATSUKA MITSUNOBU
Publication year - 1985
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1111/j.1440-169x.1985.00653.x
Subject(s) - hemicentrotus , phenylmethylsulfonyl fluoride , antipain , leupeptin , pmsf , enzyme , protease , biochemistry , intracellular , strongylocentrotus purpuratus , biology , chemistry , blastula , hatching , sea urchin , microbiology and biotechnology , embryogenesis , gastrulation , gene , zoology
The intracellular hatching enzyme was confirmed to be particulate‐bound in the sea urchin, Hemicentrotus pulcherrimus. The enzyme was solubilized most effectively by sonication in buffer containing 12.5 mM CaCl 2 , and 0.5 M KCl. The intracellular hatching enzyme is suggested to be activated by an antipain‐ or elastatinal‐susceptible protease(s) on its solubilization. Since the intracellular hatching enzyme solubilized in the absence of protease inhibitors was inhibited by phenylmethylsulfonyl fluoride (PMSF) and chymostatin, the active hatching enzyme is concluded to be a chymostatin‐sensitive serine protease. The enzyme required CaCl 2 , and KCl or NaCl for both stability and activity. The preference of the enzyme of anions as sodium salts was as follows: Cl − > NO 3 − > I − > SCN − . The apparent molecular weights of the intracellular hatching enzyme (IHE) and the hatching enzyme secreted from the blastula with or without the fertilization envelope (SHE or dSHE) were estimated as 89,000, 135,000, 80,000, respectively. On incubations with isolated fertilization envelopes as an enzyme substrate, the apparent molecular weights of dSHE and IHE increased to 128,000 and 105,000, respectively.