High molecular weight-multicatalytic proteinases in premature and mature oocytes of Rana pipiens.
High molecular weight, multicatalytic proteinases (named proteasomes) have been for the first time found, on the basis of different protein patterns, in the cytoplasmic soluble fractions of both non-hormone-treated (premature) and progesterone-treated (mature) oocytes of a frog (Rana pipiens). These enzymes, pooled separately as two fractions sedimenting between around 19S and the bottom (over 27S) on glycerol density gradient centrifugation, were composed of several molecular forms with apparent high molecular weights ranging from over 700 kDa, as judged on Sepharose 6B gel filtration. In addition, both the fractions hydrolyzed distinctly a Tyr-containing substrate in the presence of SDS as an activator, and exhibited higher activities toward Arg-containing substrates in the absence of SDS, and activity toward a Glu-containing substrate in the presence and absence of SDS. Immunological experiments using antibodies against proteasomes purified from ovaries of Xenopus laevis clearly revealed characteristic cross-reactivity with both the fractions found in Rana. These data suggest that these enzymes in the two fractions from the respective oocytes in Rana are very similar or identical to the proteasomes of Xenopus. The enzymes in premature oocytes eluted at 0.15-0.18M NaCl on a DEAE-cellulose column disappeared on treatment with TPCK, a well-known chymotrypsin inhibitor, suggesting that the 0.15-0.18M NaCl-eluate contained chymotrypsin-like proteinases probably latent in ovo. The enzymes in mature oocytes had not similar chromatographical patterns to those in premature oocytes. These results suggest that the enzymes already present in premature oocytes may be involved through conformational alterations as to the protein pattern in oocyte maturation following induction by progesterone.
Azuma
Y
Y
Department of Biology, Faculty of Science, Shizuoka University, Japan.
Tokumoto
T
T
Ishikawa
K
K
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D003593:Cytoplasm / Q000201:enzymology; D010450:Endopeptidases / Q000378:metabolism*; D005260:Female; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009865:Oocytes / Q000201:enzymology*; D011894:Rana pipiens; D012995:Solubility; D013379:Substrate Specificity; D014982:Xenopus laevis
D010450:Endopeptidases