Degradation of vitellogenins by 170 kDa trypsin-like protease in the plasma of the tilapia, Oreochromis niloticus.
Proteolytic degradation of plasma vitellogenins during purification procedure has been noted in several teleost fishes. We have characterized here a trypsin-like serine protease in the plasma of the tilapia, Oreochromis niloticus, which degrades vitellogenins. The molecular mass of the protease was estimated as 230 kDa by gel filtration and as 170 kDa both by nondenaturing and by SDS-polyacrylamide gel electrophoresis. The protease efficiently hydrolyzed the synthetic peptide substrates for trypsin-like proteases but not the substrates for chymotrypsin-like proteases nor aminopeptidases. Hydrolysis of the peptide substrates was strongly inhibited by leupeptin, aprotinin and N-tosyl-L-lysine chloromethyl ketone and to certain extent by chymostatin, 3,4-dichloroisocoumarin, phenylmethanesulfonyl fluoride, and soybean trypsin inhibitor. Leupeptin and aprotinin also inhibited the degradation of a vitellogenin in the plasma. Although the physiological functions of the 170 kDa protease in vivo have not been elucidated, the results on exzymatic properties of this protease will be useful for the isolation and characterization of vitellogenin not only in tilapia but also in other organisms.
Inaba
K
K
Misaki Marine Biological Station, Graduate School of Science, University of Tokyo, Kanagawa, Japan.
Buerano
C C
CC
Natividad
F F
FF
Morisawa
M
M
D000595:Amino Acid Sequence; D000818:Animals; D010450:Endopeptidases / Q000097:blood* / Q000737:chemistry; D004958:Estradiol / Q000494:pharmacology; D066298:In Vitro Techniques; D008297:Male; D008970:Molecular Weight; D009842:Oligopeptides / Q000737:chemistry; D011480:Protease Inhibitors / Q000494:pharmacology; D013379:Substrate Specificity; D017210:Tilapia / Q000097:blood*; D014357:Trypsin / Q000097:blood / Q000737:chemistry; D014819:Vitellogenins / Q000097:blood*
D009842:Oligopeptides; D011480:Protease Inhibitors; D014819:Vitellogenins; D004958:Estradiol; D010450:Endopeptidases; D014357:Trypsin