title: Protease activity appeared after trypsin treatment of the purified vitellogenin from eel Anguilla japonica.
abstract: Density gradient ultracentrifugation and anion-exchange chromatography combination were effective for the purification of the eel vitellogenin from the plasma of estradiol-treated eels. The vitellogenin was very high density glycolipoprotein (P = 1.27 g/ml) and its apolipoprotein was M(r) 196 k in both reduced and non-reduced conditions by SDS-PAGE. The major lipid component was phospholipid. The N-terminal amino-acid sequence of the vitellogenin was as follows: (Ac)Thr-Pro-Ala-Leu/Ala-Asp-Tyr. Amino-acid composition of the eel vitellogenin was similar to those of other teleosts. The protease activity appeared in the trypsinized vitellogenin, but was not detected in the purified vitellogenin. The protease was separated from the used trypsin and the other cleaved vitellogenin by a dextran sulfate cellulose column. The molecular weight of the protease was determined by zymogram using SDS-polyacrylamide gel containing casein and was 50 k. It was concluded that the eel vitellogenin possesses the protease activity as a latent form.
authors.affiliation: United Graduate School of Agricultural Science, Kagoshima University, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Komatsu
authors.forename: W
authors.initials: W
authors.lastname: Matsumoto
authors.forename: S
authors.initials: S
authors.lastname: Hayashi
mesh_terms: D000595:Amino Acid Sequence; D000811:Anguilla; D000818:Animals; D010450:Endopeptidases / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D004958:Estradiol / Q000494:pharmacology; D005399:Fishes; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014357:Trypsin; D014819:Vitellogenins / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D014982:Xenopus laevis
chemical_list: D010446:Peptide Fragments; D014819:Vitellogenins; D004958:Estradiol; D010450:Endopeptidases; D014357:Trypsin