Purification and characterization of maturation-promoting factor in fish

M. Yamashita, S. Fukada, Michiyasu Yoshikuni, P. Bulet, T. Hirai, Akihiko Yamaguchi, Y. H. Lou, Z. Zhao, Y. Nagahama

研究成果: ジャーナルへの寄稿記事

87 引用 (Scopus)

抄録

Maturation-promoting factor (MPF) activity has been demonstrated for the first time in fish oocytes. We purified MPF from a 100,000g supernatant of crushed, naturally spawned carp oocytes using four chromatography columns: Q-Sepharose Fast-Flow, p13sucl-affinity Sepharose, Mono S, and Superose 12. The final preparation was purified over 1000-fold with a recovery of about 1%. On Superose 12, MPF eluted as a single peak with an apparent molecular weight of 100 kDa. SDS-PAGE analysis of the active fractions after Superose 12 revealed the presence of four proteins of 33, 34, 46, and 48 kDa. A monoclonal antibody against the PSTAIR sequence of cdc2 kinase recognized the 33- and 34-kDa proteins for which the 46- and 48-kDa proteins are endogenous substrates. The 46- and 48-kDa proteins were recognized by a monoclonal antibody against Escherichia coli-produced goldfish cyclin B, but not by an anti-cyclin A antibody. When oocytes were matured in the presence of 32P, the labeling was seen with the 34-kDa protein, but not with the 33-kDa protein. The 34-kDa protein corresponded to the MPF activity, but the 33-kDa protein did not. These findings indicate that carp MPF is a complex of cdc2 kinase and cyclin B, and further that active MPF contains the phosphorylated form of cdc2 kinase.

元の言語英語
ページ(範囲)8-15
ページ数8
ジャーナルDevelopmental Biology
149
発行部数1
DOI
出版物ステータス出版済み - 1 1 1992

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Maturation-Promoting Factor
Fishes
Proteins
Oocytes
Cyclin B
Phosphotransferases
Carps
Sepharose
Monoclonal Antibodies
Cyclin A
Goldfish
Chromatography
Polyacrylamide Gel Electrophoresis
Molecular Weight
Escherichia coli

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

これを引用

Purification and characterization of maturation-promoting factor in fish. / Yamashita, M.; Fukada, S.; Yoshikuni, Michiyasu; Bulet, P.; Hirai, T.; Yamaguchi, Akihiko; Lou, Y. H.; Zhao, Z.; Nagahama, Y.

:: Developmental Biology, 巻 149, 番号 1, 01.01.1992, p. 8-15.

研究成果: ジャーナルへの寄稿記事

Yamashita, M, Fukada, S, Yoshikuni, M, Bulet, P, Hirai, T, Yamaguchi, A, Lou, YH, Zhao, Z & Nagahama, Y 1992, 'Purification and characterization of maturation-promoting factor in fish', Developmental Biology, 巻. 149, 番号 1, pp. 8-15. https://doi.org/10.1016/0012-1606(92)90259-J
Yamashita, M. ; Fukada, S. ; Yoshikuni, Michiyasu ; Bulet, P. ; Hirai, T. ; Yamaguchi, Akihiko ; Lou, Y. H. ; Zhao, Z. ; Nagahama, Y. / Purification and characterization of maturation-promoting factor in fish. :: Developmental Biology. 1992 ; 巻 149, 番号 1. pp. 8-15.
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abstract = "Maturation-promoting factor (MPF) activity has been demonstrated for the first time in fish oocytes. We purified MPF from a 100,000g supernatant of crushed, naturally spawned carp oocytes using four chromatography columns: Q-Sepharose Fast-Flow, p13sucl-affinity Sepharose, Mono S, and Superose 12. The final preparation was purified over 1000-fold with a recovery of about 1{\%}. On Superose 12, MPF eluted as a single peak with an apparent molecular weight of 100 kDa. SDS-PAGE analysis of the active fractions after Superose 12 revealed the presence of four proteins of 33, 34, 46, and 48 kDa. A monoclonal antibody against the PSTAIR sequence of cdc2 kinase recognized the 33- and 34-kDa proteins for which the 46- and 48-kDa proteins are endogenous substrates. The 46- and 48-kDa proteins were recognized by a monoclonal antibody against Escherichia coli-produced goldfish cyclin B, but not by an anti-cyclin A antibody. When oocytes were matured in the presence of 32P, the labeling was seen with the 34-kDa protein, but not with the 33-kDa protein. The 34-kDa protein corresponded to the MPF activity, but the 33-kDa protein did not. These findings indicate that carp MPF is a complex of cdc2 kinase and cyclin B, and further that active MPF contains the phosphorylated form of cdc2 kinase.",
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