Trypsin-like hatching protease from mouse embryos: evidence for the presence in culture medium and its enzymatic properties.
Enzymatic properties of a protease involved in hatching of mouse embryos were examined. A trypsin-like protease, which most efficiently hydrolyzed t-butoxycarbonyl-Leu-Ser-Thr-Arg-4-methylcoumaryl-7-amide, was demonstrated in culture medium of mouse hatching embryos. The enzyme was strongly inhibited by diisopropyl fluorophosphate, phenylmethanesulfonyl fluoride, leupeptin, antipain, N alpha-tosyl-L-lysyl-chloromethane, soybean trypsin inhibitor, and Trasylol, but not or weakly inhibited by p-chloromercuribenzoic acid, EDTA, E-64, pepstatin, chymostatin, and bestatin, suggesting a trypsin-like serine proteinase. The protease activity in the medium gradually elevated during the course of hatching, whereas the embryo-associated activity showed no significant change. Furthermore, pyroglutamyl-Leu-argininal, the strongest inhibitor for the enzyme among peptidyl argininals, all of which are potent trypsin inhibitors, showed the strongest inhibition toward hatching. Thus, a trypsin-like protease secreted from hatching embryos into the culture medium may participate in mouse hatching, probably as a hatching enzyme.
Sawada
H
H
Department of Life Science, Faculty of Science, Tokyo Institute of Technology, Japan.
Yamazaki
K
K
Hoshi
M
M
D000818:Animals; D001755:Blastocyst / Q000187:drug effects; D003374:Coumarins / Q000378:metabolism; D003470:Culture Media / Q000378:metabolism; D004622:Embryo, Mammalian / Q000201:enzymology*; D007976:Leupeptins / Q000494:pharmacology; D008666:Metalloendopeptidases / Q000032:analysis* / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D009842:Oligopeptides / Q000378:metabolism; D010447:Peptide Hydrolases / Q000032:analysis* / Q000378:metabolism; D011480:Protease Inhibitors; D012697:Serine Endopeptidases / Q000032:analysis* / Q000378:metabolism; D013379:Substrate Specificity
D003374:Coumarins; D003470:Culture Media; D007976:Leupeptins; D009842:Oligopeptides; D011480:Protease Inhibitors; C033868:Boc-leucyl-seryl-threonyl-arginine-4-methylcoumaryl-7-amide; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases; C048419:strypsin; D008666:Metalloendopeptidases; C024344:envelysin; C032854:leupeptin