1990 Fiscal Year Annual Research Report

蛋白質チロシン残基の燐酸化とその細胞生物学的意義

Research Project

Project/Area Number	02304034
Research Institution	福井医科大学
Principal Investigator	山村博平福井医科大学, 医学部, 教授 (90030882)
Co-Investigator(Kenkyū-buntansha)	山本雅東京大学, 医科研, 助教授 (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)
Keywords	p60^<cーsrc> / p60^<cーyes> / PDGF受容体 / インスリン受容体 / PIキナ-ゼ / EGF受容体 / 蛋白質チロシンホスファタ-ゼ / 細胞質チロシンキナ-ゼ
Research Abstract	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ーsrcの負の活性調節部位であるTyrー527を特異的に燐酸化する極めてユニ-クなチロシンキナ-ゼ(cーsrc kinase)のcDNAクロ-ニングに成功した。2)ラット肝形質膜よりp60cーyesの部分精製に成功し自己燐酸化によって活性が著しく上昇することを明らかにした。3)神経成長円錐、形質転換あるいは癌化細胞の基質接着部などの動的接着部に於けるsrc遺伝子産物の集積を見いだした。4)cーraf蛋白がインスリン受容体、PDGF受容体によって直接チロシン残基が燐酸化されれないことを明らかにした。5)インスリン受容体が新しいタイプのPIキナ-ゼであるPIー3キナ-ゼの85Kサブユニットを燐酸化することを明らかにした。6)インスリン受容体異常症タイプA(山梨)においてインスリン受容体遺伝子のエクソン14の欠失を明かとした。7)ヒト上皮癌由来のA431細胞において、細胞内cAMPは少なくともCキナ-ゼの活性化因子であるジアシルグリセロ-ルの産生を阻害することによってEGF受容体シグナル変換系の出力を抑制することを示した。8)Tリンパ球においてFyn蛋白質がTCRーCD3複合体とまたBリンパ球においては、Lyn蛋白質がmIgMやmIgDと物理的に会合していることを示した。9)ブタ脾臓の細胞質チロシンキナ-ゼの活性制御には酵素活性に影響を与える因子の基質側の修飾も重要であることを明らかにし、生理的基質を検討し細胞質、ミクロゾ-ム、核画分に多数存在することを明かにした。10)ラット肝臓可溶画分より2種類の蛋白質チロシンホスファタ-ゼ、PTPー1,PTPー2を精製した。このうちPTPー1はこれまでに報告のない新しいタイプの蛋白質チロシンホスファタ-ゼであることを明らかにした。

Research Products

(23 results)

All Other

All Publications (23 results)

[Publications] Hashimoto,Eikichi: "Comparison of substrate recognition by protein kinase (type III) between rat liver cytosolic and particulate fractions." Int.J.Biochem.22(4). 405-410 (1990)
[Publications] Kobayashi,Tomoko: "Purification and characterization of a cytosolic proteinーtyrosine kinase from porcine spleen." Eur.J.Biochem.188(3). 535-540 (1990)
[Publications] Asahi,Momoyo: "Diverse effects of polyーbasic amino acids,heparin and ionic strength on the phosphorylation of various substrates by cytosolic proteinーtyrosine kinase from porcine spleen." Int.J.Biochem.22(6). 635-640 (1990)
[Publications] Inazu,Tetuya: "Proteinーtyrosine phosphorylation and aggregation of intact human platelets by vanadate with H_2O_2." Biochem.Biophys.Res.Commun.170(1). 259-263 (1990)
[Publications] Hashimoto,Eikichi: "Mechanism of proteolytic activation of rat liver protein kinase C generating Ca^<2+>ー phospholipidーindependent form with apparent molecular mass of 80,000." Int.J.Biochem.22(8). 865-871 (1990)
[Publications] Hashimoto,Eikichi: "Proteaseーactivated form of protein kinase C with Mr 80,000 generated from rat liver plasma membrane by trypsinーlike protease." Biochem.Int.21(5). 949-957 (1990)
[Publications] Yamamoto,R.: "Phosphorylation state and biological function of a mutant human insulin receptor." J.Biol.Chem.265. 14777-14783 (1990)
[Publications] Shimada F.: "Insulin resistant diabetes associated with partial deletion of insulin receptor gene" Lancet. 335. 1179-1181 (1990)
[Publications] Araki E.: "A cluster of four Spl binding sites required for efficient expression of the human insulin receptor gene" J.Biol.Chem.
[Publications] Azuma,Kazuyoshi: "Purification and characterization of a rat liver membrane tyrosineーprotein kinase,the possible protooncogene cーyes product,p60^<cーyes>." J.Biol.Chem.
[Publications] Nakata,T.: "Conformational change and localization of calpactin I complex involved in exocytosis as revealed by quickーfreeze,deepーetch,electron microscopy and immunocytochemistry." J.Cell Biol.110. 13-25 (1990)
[Publications] Sobue.K.: "Involvement of the membrane cytoskeletal proteins and the src gene product in growth cone adhesion and movement." Neuroscience Res.Suppl.13. 80-91 (1990)
[Publications] Hashimoto,Eikichi: "Studies on protein kinase C tightlyーbound to rat liver plasma membrane and its proteaseーactivated form." Int.J.Biochem.23(4). 395-403 (1990)
[Publications] Hashimoto,Eikichi: "Proteaseーactivated protein kinase C in rat liver." Int.J.Biochem.
[Publications] Shimomura,Reiko: "Possible endogenous substrate proteins for cytosolic proteinーtyrosine kinase from porcine spleen." Int.J.Biochem.
[Publications] Takayama,Hiroshi: "Ionophore A23187ーinduced proteinーtyrosine phosphorylation of human platelets:Possible synergism between Ca^<2+> mobilization and protein kinase C activation." Biochem.Biophys.Res.Commun.
[Publications] Inazu,Tetuya: "The lectin wheat germ agglutinin induces rapid proteinーtyrosine phosphorylation in human platelets." Biochem.Biophys.Res.Commun.
[Publications] Nada,S.: "Molecular cloning of a proteinーtyrosine kinase that specifically phosphorylates a negativeーregulatory site of p60^<cーsrc>." Nature.
[Publications] Inoue,Kazushi: "Specific expression of human cー<fgr>___ー in monocytes and large granular lymphocytes,effectors of natural immunity." Mol.Cell.Biol.10. 1789-1792 (1990)
[Publications] Yagi,Takeshi: "Homologous recombination at cーfyn locus of mous embryonig stem cells with diphtheria toxin A fragment gene in negative selection." Proc Natl.Acad.Sci.
[Publications] Iwashita,Shintarou: "cAMPーmediated Modulation of signal transduction of EGF receptor systems in human epidermoid carcinoma A431 cells." J.Biol.Chem.265. 10702-10708 (1990)
[Publications] Iwashita,Shintarou: "Examination of the role of serine phosphorylation in PLCーg and its related p47 in cAMPーmediated depression of EGF receptor signal transduction." FEBS Letters. 268. 157-160 (1990)
[Publications] 立木蔚: "蛋白質チロシンホスファタ-ゼとその役割" 代謝. 27(9). 93-101 (1990)

1990 Fiscal Year Annual Research Report

蛋白質チロシン残基の燐酸化とその細胞生物学的意義

Principal Investigator

山村 博平 福井医科大学, 医学部, 教授 (90030882)

Research Products

[Publications] Hashimoto,Eikichi: "Comparison of substrate recognition by protein kinase (type III) between rat liver cytosolic and particulate fractions." Int.J.Biochem.22(4). 405-410 (1990)

[Publications] Kobayashi,Tomoko: "Purification and characterization of a cytosolic proteinーtyrosine kinase from porcine spleen." Eur.J.Biochem.188(3). 535-540 (1990)

[Publications] Asahi,Momoyo: "Diverse effects of polyーbasic amino acids,heparin and ionic strength on the phosphorylation of various substrates by cytosolic proteinーtyrosine kinase from porcine spleen." Int.J.Biochem.22(6). 635-640 (1990)

[Publications] Inazu,Tetuya: "Proteinーtyrosine phosphorylation and aggregation of intact human platelets by vanadate with H_2O_2." Biochem.Biophys.Res.Commun.170(1). 259-263 (1990)

[Publications] Hashimoto,Eikichi: "Mechanism of proteolytic activation of rat liver protein kinase C generating Ca^<2+>ー phospholipidーindependent form with apparent molecular mass of 80,000." Int.J.Biochem.22(8). 865-871 (1990)

[Publications] Hashimoto,Eikichi: "Proteaseーactivated form of protein kinase C with Mr 80,000 generated from rat liver plasma membrane by trypsinーlike protease." Biochem.Int.21(5). 949-957 (1990)

[Publications] Yamamoto,R.: "Phosphorylation state and biological function of a mutant human insulin receptor." J.Biol.Chem.265. 14777-14783 (1990)

[Publications] Shimada F.: "Insulin resistant diabetes associated with partial deletion of insulin receptor gene" Lancet. 335. 1179-1181 (1990)

[Publications] Araki E.: "A cluster of four Spl binding sites required for efficient expression of the human insulin receptor gene" J.Biol.Chem.

[Publications] Azuma,Kazuyoshi: "Purification and characterization of a rat liver membrane tyrosineーprotein kinase,the possible protooncogene cーyes product,p60^<cーyes>." J.Biol.Chem.

[Publications] Nakata,T.: "Conformational change and localization of calpactin I complex involved in exocytosis as revealed by quickーfreeze,deepーetch,electron microscopy and immunocytochemistry." J.Cell Biol.110. 13-25 (1990)

[Publications] Sobue.K.: "Involvement of the membrane cytoskeletal proteins and the src gene product in growth cone adhesion and movement." Neuroscience Res.Suppl.13. 80-91 (1990)

[Publications] Hashimoto,Eikichi: "Studies on protein kinase C tightlyーbound to rat liver plasma membrane and its proteaseーactivated form." Int.J.Biochem.23(4). 395-403 (1990)

[Publications] Hashimoto,Eikichi: "Proteaseーactivated protein kinase C in rat liver." Int.J.Biochem.

[Publications] Shimomura,Reiko: "Possible endogenous substrate proteins for cytosolic proteinーtyrosine kinase from porcine spleen." Int.J.Biochem.

[Publications] Takayama,Hiroshi: "Ionophore A23187ーinduced proteinーtyrosine phosphorylation of human platelets:Possible synergism between Ca^<2+> mobilization and protein kinase C activation." Biochem.Biophys.Res.Commun.

[Publications] Inazu,Tetuya: "The lectin wheat germ agglutinin induces rapid proteinーtyrosine phosphorylation in human platelets." Biochem.Biophys.Res.Commun.

[Publications] Nada,S.: "Molecular cloning of a proteinーtyrosine kinase that specifically phosphorylates a negativeーregulatory site of p60^<cーsrc>." Nature.

[Publications] Inoue,Kazushi: "Specific expression of human cー<fgr>___ー in monocytes and large granular lymphocytes,effectors of natural immunity." Mol.Cell.Biol.10. 1789-1792 (1990)

[Publications] Yagi,Takeshi: "Homologous recombination at cーfyn locus of mous embryonig stem cells with diphtheria toxin A fragment gene in negative selection." Proc Natl.Acad.Sci.

[Publications] Iwashita,Shintarou: "cAMPーmediated Modulation of signal transduction of EGF receptor systems in human epidermoid carcinoma A431 cells." J.Biol.Chem.265. 10702-10708 (1990)

[Publications] Iwashita,Shintarou: "Examination of the role of serine phosphorylation in PLCーg and its related p47 in cAMPーmediated depression of EGF receptor signal transduction." FEBS Letters. 268. 157-160 (1990)

[Publications] 立木 蔚: "蛋白質チロシンホスファタ-ゼとその役割" 代謝. 27(9). 93-101 (1990)

山村博平福井医科大学, 医学部, 教授 (90030882)

[Publications] 立木蔚: "蛋白質チロシンホスファタ-ゼとその役割" 代謝. 27(9). 93-101 (1990)