1991 年度実績報告書

蛋白質チロシン残基の燐酸化とその細胞生物学的意義

研究課題

研究課題/領域番号	02304034
研究機関	福井医科大学
研究代表者	山村博平福井医科大学, 医学部, 教授 (90030882)
研究分担者	山本雅東京大学, 医科研, 教授 (40134621) 蛯名洋介徳島大学, 酵素研, 教授 (00112227) 武田誠郎広島大学, 医学部, 教授 (40030853) 春日雅人神戸大学, 医学部, 教授 (50161047) 中川八郎大阪大学, 蛋白研, 教授 (20029937)
キーワード	CDNAクロ-ニング / CSK / p60^<src> / インスリン受容体 / PI‐3キナ-ゼ / EGF受容体チロシンキナ-ゼ / p72^<syk> / 蛋白質チロシンホスファタ-ゼI
研究概要	蛋白質チロシン残基の燐酸化の細胞生物学的意義に関して今年度は以下の点について明らかにした。発癌遺伝子産物に関して1)p60c‐srcの負の活性調節部位(Tyu‐527)を特異的に燐酸化するチロシンキナ-ゼ(csk)のcDNAクロ-ニングに成功し、cskがsrc‐familyチロシンキナ-ゼ以外の活性制御にもかかわる可能性を示した。2)ラット脳顆粒画分より癌原遺伝子c‐yes,c‐src,fynの産物であるsrc‐familyチロシンキナ-ゼを分離精製した。p59fynは電気泳動的に単一バンドにまで精製した。精製したp59fynにチロシン残基の自己燐酸化がみられた。3)p60srcの基質となるアネキシン2と6の細胞局在について検討を行なった結果、src感染細胞の動的接着部と正常細胞の基質接着部にアネキシン6が局在し、動的接着部のみにアネキシン2が分布することを見いだした。4)TypeCインスリン受容体異常症(千葉)のEBウイルスでトランスフォ-ムしたリンパ球を用い、異常インスリン受容体mRNA及び蛋白質を同定した。5)Site‐directed mutagenesisの手法を用いて各種のインスリン受容体mutantを作製してインスリン受容体の構造と機能の関係を明らかにした。6)In vitroにおいてPI‐3キナ-ゼ自身またはそれにassociateする蛋白質がインスリン受容体チロシンキナ-ゼにより燐酸化されPI‐3キナ-ゼの活性が高まる可能性を示唆した。7)EGF受容体チロシンキナ-ゼの標的蛋白質の1つGTPase activating Protein(GAP)の比活性が細胞増殖の静止期では対数増殖期の場合に比べて約2倍上昇していることを明らかにした。8)Bリンパ球の抗原受容体を刺激することにより1ynが活性化されることまたTリンパ球にfynを過剰発現させることにより、抗原受容体を介したTリンパ球活性化反応が促進されることを明らかにした。9)ブタ脾臓より72kDaのチロシンキナ-ゼのクロ-ニングに成功した。この新しいチロシンキナ-ゼの遺伝子をsykと名付けた。p72sykはWGAによって自己燐酸化反応が促進されることを明らかにした。10)ラット肝蛋白質チロシンホスファタ-ゼ‐1(PTPase‐1)の部分アミノ酸配列解析の結果、PTPase‐1は2つのsrcホモロジー領域ー2(SHー2)と1つの解媒ドメインをもつことを明らかにした。

研究成果

(28件)

すべてその他

すべて文献書誌 (28件)

[文献書誌] Eikichi Hashimoto: "Studies on protein kinase C tightly‐bound to rat liver plasma membrane and its protease‐activated form." Int.J.Biochem.24(4). 395-403 (1991)
[文献書誌] Eikichi Hashimoto: "Protease‐activated protein kinase C in rat liver." Int.J.Biochem.23(5,6). 507-512 (1991)
[文献書誌] Reiko Shimomura: "Possible endogenous substrate proteins for cytosolic protein‐tyrosine kinase from porcine spleen." Int.J.Biochem.23(7,8). 727-731 (1991)
[文献書誌] Hiroshi Takayama: "Ionophore A23187‐induced protein‐tyrosine phosphorylation of human platelets:Possible synergism between Ca^<2+>mobilization and protein kinase C activation." Biochem.Biophys.Res.Commun.174(2). 992-997 (1991)
[文献書誌] Tetsuya Inazu: "The lectin wheat germ agglutinin induces rapid protein‐tyrosine phosphorylation in human platelets." Biochem.Biophys.Res.Commun.174(3). 1154-1158 (1991)
[文献書誌] Takanobu Taniguchi: "Molecular cloning of a porcine gene syk that encodes a 72‐kDa protein‐tyrosine kinase showing high susceptibility to proteolysis." J.Biol.chem.266(24). 15790-15796 (1991)
[文献書誌] Takechiyo Yamada: "The lectin wheat germ agglutinin stimulates a protein‐tyrosine kinase activity of p72^<syk> in porcine splenocytes." Biochem.Biophys.Res.Commun.180(3). 1325-1329 (1991)
[文献書誌] Keiko Sakai: "Identification of the phosphorylation sites of H2B histone by a catalytic fragment of p72^<syk> from porcine spleen." FEBS Letters. 294(1,2). 104-108 (1991)
[文献書誌] Fumito Takeuchi: "Phosphorylation of Histone H2A by protein kinase C and Identification of the Phosphorylation Site." J.Biochem.
[文献書誌] Nada,S.: "Cloning of a complementary DNA for a protein‐tyrosine kinase that specifically phosphorylates a negative regulatory site of p60^<cーsrc>" Nature. 351. 69-72 (1991)
[文献書誌] Okada,M.: "CSK:A protein‐tyrosine Kinase involved in regulation of src family kinases" J.Biol.Chem.
[文献書誌] Tobe,K.: "Insulin and 12‐o‐Tetradecanoyl phorbol 13‐Acetate(TPA)Activation of Two lmmunologically Distinct MBP/MAP2 Kinases via de novo Phosphorylation of Threonine and Tyrosine Residues" J.Biol.Chem.
[文献書誌] Yonezawa,K.: "In Vitro Association of Phosphatidylinositol 3‐Kinase Activity with Activated Insulin Receptor Tyrosine Kinase" J.Biol.Chem.
[文献書誌] Kazuyoshi Azuma: "Purification and characterization of a rat liver membrane tyrosine‐protein kinase, the possible protooncogene c‐yes product,p60^<c-yes>." J.Biol.Chem.266. 4831-4839 (1991)
[文献書誌] Hideki H.: "Phosphorylation in vitro of the 85 kDa subunit of phosphatidylinositol 3‐kinase and its possible activation by insulin receptor tyrosine kinase" Biochem.J.
[文献書誌] Suzuki,Y.: "Defects in insulin binding and receptor kinase in cells from a woman with type A insulin resistance and from her family." Diabetologia. 34. 86-92 (1991)
[文献書誌] Tokuyama,Y: "Hypersecretion of islet amyloid polypeptide from pancreatic islets of ventromedial hypothalamic‐lesioned rats and obese Zucker rats." Endocrinology. 128. 2739-2744 (1991)
[文献書誌] Yuji Yamanashi: "Associations of B cell antigen receptor and protein‐tyrosine kinase p50^<lyn>." Science. 251. 192-194 (1991)
[文献書誌] Yuji Yamanashi: "Differential responses of p56^<lyn> and p53^<lyn>,products of alternatively spliced lynmRNA,upon stimulation of B cell antigen receptor." Cell Regulation.
[文献書誌] Hiraga,A.: "Rat liver cytosolic protein‐tyrosine phosphatases:Purification and characterization of two forms." Adv.Prptein Phosphatases. 6. 251-264 (1991)
[文献書誌] Shintaro Iwashita: "Signal transduction system for growth factor receptors associated with tyrosine kinase activity‐Epidermal growth factor receptor signalling and its regulation" Cellular Signalling.
[文献書誌] T.Yamori: "Insulin‐like growth factor I rapidly induces tyrosine phosphorylation of a Mr150.000 and a Mr160.000 protein in highly metastatic mouse colon carcinoma 26NL‐17 cells" Cancer Research.
[文献書誌] Sobue K.: "Caldesmon,a Novel Regulatory Protein in Smooth Muscle and Nonmuscle Actomyosin Systems" J.Biol.Chem.266. 12115-12118 (1991)
[文献書誌] Tanaka T.: "Ca^<2+>‐dependent Regulation of the Spectrin/Actin Interaction by Calmodulin and Protein 4.1" J.Biol.Chem.266. 1134-1140 (1991)
[文献書誌] 山村博平: "病理学キ-ワ-ド" 文光堂, 1 (1991)
[文献書誌] 山村博平: "病理学キ-ワ-ド" 文光堂, 1 (1991)
[文献書誌] 稲津哲也: "代謝" 中山書店, 5 (1991)
[文献書誌] 谷口隆信: "注目の臨床実験検査法" 中山書店, 9 (1991)

1991 年度 実績報告書

蛋白質チロシン残基の燐酸化とその細胞生物学的意義

研究代表者

山村 博平 福井医科大学, 医学部, 教授 (90030882)

研究成果

[文献書誌] Eikichi Hashimoto: "Studies on protein kinase C tightly‐bound to rat liver plasma membrane and its protease‐activated form." Int.J.Biochem.24(4). 395-403 (1991)

[文献書誌] Eikichi Hashimoto: "Protease‐activated protein kinase C in rat liver." Int.J.Biochem.23(5,6). 507-512 (1991)

[文献書誌] Reiko Shimomura: "Possible endogenous substrate proteins for cytosolic protein‐tyrosine kinase from porcine spleen." Int.J.Biochem.23(7,8). 727-731 (1991)

[文献書誌] Hiroshi Takayama: "Ionophore A23187‐induced protein‐tyrosine phosphorylation of human platelets:Possible synergism between Ca^<2+>mobilization and protein kinase C activation." Biochem.Biophys.Res.Commun.174(2). 992-997 (1991)

[文献書誌] Tetsuya Inazu: "The lectin wheat germ agglutinin induces rapid protein‐tyrosine phosphorylation in human platelets." Biochem.Biophys.Res.Commun.174(3). 1154-1158 (1991)

[文献書誌] Takanobu Taniguchi: "Molecular cloning of a porcine gene syk that encodes a 72‐kDa protein‐tyrosine kinase showing high susceptibility to proteolysis." J.Biol.chem.266(24). 15790-15796 (1991)

[文献書誌] Takechiyo Yamada: "The lectin wheat germ agglutinin stimulates a protein‐tyrosine kinase activity of p72^<syk> in porcine splenocytes." Biochem.Biophys.Res.Commun.180(3). 1325-1329 (1991)

[文献書誌] Keiko Sakai: "Identification of the phosphorylation sites of H2B histone by a catalytic fragment of p72^<syk> from porcine spleen." FEBS Letters. 294(1,2). 104-108 (1991)

[文献書誌] Fumito Takeuchi: "Phosphorylation of Histone H2A by protein kinase C and Identification of the Phosphorylation Site." J.Biochem.

[文献書誌] Nada,S.: "Cloning of a complementary DNA for a protein‐tyrosine kinase that specifically phosphorylates a negative regulatory site of p60^<cーsrc>" Nature. 351. 69-72 (1991)

[文献書誌] Okada,M.: "CSK:A protein‐tyrosine Kinase involved in regulation of src family kinases" J.Biol.Chem.

[文献書誌] Tobe,K.: "Insulin and 12‐o‐Tetradecanoyl phorbol 13‐Acetate(TPA)Activation of Two lmmunologically Distinct MBP/MAP2 Kinases via de novo Phosphorylation of Threonine and Tyrosine Residues" J.Biol.Chem.

[文献書誌] Yonezawa,K.: "In Vitro Association of Phosphatidylinositol 3‐Kinase Activity with Activated Insulin Receptor Tyrosine Kinase" J.Biol.Chem.

[文献書誌] Kazuyoshi Azuma: "Purification and characterization of a rat liver membrane tyrosine‐protein kinase, the possible protooncogene c‐yes product,p60^<c-yes>." J.Biol.Chem.266. 4831-4839 (1991)

[文献書誌] Hideki H.: "Phosphorylation in vitro of the 85 kDa subunit of phosphatidylinositol 3‐kinase and its possible activation by insulin receptor tyrosine kinase" Biochem.J.

[文献書誌] Suzuki,Y.: "Defects in insulin binding and receptor kinase in cells from a woman with type A insulin resistance and from her family." Diabetologia. 34. 86-92 (1991)

[文献書誌] Tokuyama,Y: "Hypersecretion of islet amyloid polypeptide from pancreatic islets of ventromedial hypothalamic‐lesioned rats and obese Zucker rats." Endocrinology. 128. 2739-2744 (1991)

[文献書誌] Yuji Yamanashi: "Associations of B cell antigen receptor and protein‐tyrosine kinase p50^<lyn>." Science. 251. 192-194 (1991)

[文献書誌] Yuji Yamanashi: "Differential responses of p56^<lyn> and p53^<lyn>,products of alternatively spliced lynmRNA,upon stimulation of B cell antigen receptor." Cell Regulation.

[文献書誌] Hiraga,A.: "Rat liver cytosolic protein‐tyrosine phosphatases:Purification and characterization of two forms." Adv.Prptein Phosphatases. 6. 251-264 (1991)

[文献書誌] Shintaro Iwashita: "Signal transduction system for growth factor receptors associated with tyrosine kinase activity‐Epidermal growth factor receptor signalling and its regulation" Cellular Signalling.

[文献書誌] T.Yamori: "Insulin‐like growth factor I rapidly induces tyrosine phosphorylation of a Mr150.000 and a Mr160.000 protein in highly metastatic mouse colon carcinoma 26NL‐17 cells" Cancer Research.

[文献書誌] Sobue K.: "Caldesmon,a Novel Regulatory Protein in Smooth Muscle and Nonmuscle Actomyosin Systems" J.Biol.Chem.266. 12115-12118 (1991)

[文献書誌] Tanaka T.: "Ca^<2+>‐dependent Regulation of the Spectrin/Actin Interaction by Calmodulin and Protein 4.1" J.Biol.Chem.266. 1134-1140 (1991)

[文献書誌] 山村 博平: "病理学キ-ワ-ド" 文光堂, 1 (1991)

[文献書誌] 山村 博平: "病理学キ-ワ-ド" 文光堂, 1 (1991)

[文献書誌] 稲津 哲也: "代謝" 中山書店, 5 (1991)

[文献書誌] 谷口 隆信: "注目の臨床実験検査法" 中山書店, 9 (1991)

1991 年度実績報告書

山村博平福井医科大学, 医学部, 教授 (90030882)

[文献書誌] 山村博平: "病理学キ-ワ-ド" 文光堂, 1 (1991)

[文献書誌] 山村博平: "病理学キ-ワ-ド" 文光堂, 1 (1991)

[文献書誌] 稲津哲也: "代謝" 中山書店, 5 (1991)

[文献書誌] 谷口隆信: "注目の臨床実験検査法" 中山書店, 9 (1991)