1990 年度実績報告書

低分子量GTP結合蛋白質を介する新しい細胞内情報伝達機構

研究課題

研究課題/領域番号	01480529
研究機関	神戸大学
研究代表者	高井義美神戸大学, 医学部, 教授 (60093514)
研究分担者	河田正仁神戸大学, 医学部, 助手 (20224785) 貝淵弘三神戸大学, 医学部, 助教授 (00169377)
キーワード	低分子量GTP結合蛋白質 / GDP解離促進蛋白質 / GDP解離抑制蛋白質 / ゲニラルゲラニル基 / クロスト-ク / GTPase活性促進蛋白質
研究概要	昨年度の本研究において、私共は低分子量G蛋白質の活性制御蛋白質として、<smg>___ーp21と<rho>___ーp21に対するGTPase活性促進蛋白質(GAP)および<smg>___ーp25Aと<rho>___ーp21に対するGDP解離抑制蛋白質(GDI)を見出した。本年度の研究においてまず、私共はGAPやGDIとは異なる活性制御蛋白質であるGDP解離促進蛋白質(GDS)を<smg>___ーp21と<rho>___ーp21に対して見出し、<smg>___ーp21GDSのcDNAを単離して、その一次構造を決定した。また、私共は低分子量G蛋白質は細胞膜へ結合するが、GDIとGDSはこの低分子量G蛋白質の細胞膜への結合を制御することを見出した。次に、私共はC末端側のアミノ酸配列がCysーAーAーXである<smg>___ーp21Bと<rho>___ーAp21の翻訳後修飾を解析して、両低分子量G蛋白質のC末端側のCys残基には<ras>___ーp21のファルネシル基とは異なるゲラニルゲラニル基が結合し、AーAーX部分は取りのぞかれ、Cys残基がメチル化を受けることを明らかにした。また、<smg>___ーp21AのC末端はCysーXーCysと<ras>___ーp21とは異なっているが、少くとも一方のCys残基にはゲラニルゲラニル基が結合していることも明らかにした。上述したように、これらの低分子量G蛋白質は細胞膜へ結合したが、翻訳後修飾機構のない大腸菌で産生された低分子量G蛋白質は細胞膜へは結合しなかった。したがって、これらの翻訳後修飾は低分子量G蛋白質の細胞内局在や機能発現に重要であると考えられた。続いて、低分子量G蛋白質の機能を解析して<smg>___ーp21が<ras>___ーp21GAP活性を抑制することを明らかにした。したがって、<smg>___ーp21は<ras>___ーp21GAP活性を抑制することにより<ras>___ーp21の作用を促進することが示唆された。私共はすでに<smg>___ーp21がAキナ-ゼによりリン酸化されることを報告しているので、<ras>___ーp21がチロシンキナ-ゼ系の下流に位置するという最近の報告とあわせて<smg>___ーp21はチロキシンキナ-ゼ系とAキナ-ゼ系のクロスト-クに重要であると考えられた。以上の成績を得たことにより、本年度の研究計画はほぼ達成することができたと考えている。

研究成果

(36件)

すべてその他

すべて文献書誌 (36件)

[文献書誌] Fukumoto,Y.: "Activation of the cーfos serumーresponse element by the activated cーHaーras protein in a manner independent of protein kinase C and cAMPーdependent protein kinase." J.Biol.Chem.265. 774-780 (1990)
[文献書誌] Sasaki,T.: "Purification and characterization from bovine cytosol of a protein that inhibits the dissociation of GDP from and the subsequent binding of GTP to smg p25A,a ras p21ーlike GTPーbinding protein." J.Biol.Chem.265. 2333-2337 (1990)
[文献書誌] Hata,Y.: "Inhibition of the ras p21 GTPaseーactivating proteinーstimulated GTPase activity of cーHaーras p21 by smg p21 having the same putative effector domain as ras p21s." J.Biol.Chem.265. 7104-7107 (1990)
[文献書誌] Ueda,T.: "Purification and characterization from bovine brain cytosol of a novel regulatory protein inhibiting the dissociation of GDP from and the subsequent binding of GTP to rhoB p20,a ras p21ーlike GTPーbinding protein." J.Biol.Chem.265. 9373-9380 (1990)
[文献書誌] Mizoguchi,A.: "Localization and subcellular distribution of smg p25A,a ras p21ーlike GTPーbinding protein,in rat brain." J.Biol.Chem.265. 11872-11879 (1990)
[文献書誌] Araki,S.: "Regulation of reversible binding of smg p25A,a ras p21ーlike GTPーbinding protein,to synaptic plasma membranes and vesicles by its regulatory protein,GDP dissociation inhibitor." J.Biol.Chem.265. 13007-13015 (1990)
[文献書誌] Yamamoto,T.: "Purification and characterization from bovine brain cytosol of proteins that regulate the GDP/GTP exchange reaction of smg p21s,ras p21ーlike GTPーbinding proteins." J.Biol.Chem.265. 16626-16634 (1990)
[文献書誌] Kim,S.: "Tissue and subcellular distributions of the smg 21/rap1/Krevー1 proteins which are partly distinct from those of cーras p21s" Mol.Cell.Biol.10. 2645-2652 (1990)
[文献書誌] Matsui,Y.: "Molecular cloning and characterization of a novel type of regulatory protein (GDI) for smg p25A,a ras p21ーlike GTPーbinding protein." Mol.Cell.Biol.10. 4116-4122 (1990)
[文献書誌] Hoshijima,M.: "Purfication and characterization from bovine brain membranes of a GTPーbinding protein with a Mr of 21,000 ADPーribosylated by an ADPーribosyltransferase contaminated in botulinum toxin type C1ーidentification as the rhoA gene product." Mol.Brain Res.7. 9-16 (1990)
[文献書誌] Yamamoto,J.: "A GtPaseーactivating protein for rhoB p20,a ras p21ーlike GTPーbinding proteinーpartial purification,characterization and subcellular distribution in rat brain." Mol.Brain Res.8. 105-111 (1990)
[文献書誌] Sano,K.: "Differential expreession of the Nーmyc,cーfos,and smg p25A genes in human neuroblastoma cells during neuronal and Schwannian differentiation." Mol.Brain Res.9. 149-152 (1990)
[文献書誌] Kawata,M.: "Posttranslationally processed structure of the human platelet protein smg p21B:Evidence for geranylgeranylation and carboxyl methylation of the Cーterminal cysteine." Proc.Natl.Acad.Sci.USA. 87. 8960-8964 (1990)
[文献書誌] Sano,K.: "Expression of the smg p25A (a ras p21ーlike GTPーbinding protein) gene in human neuroblastoma cell lines and tumor tissues." Canser Res.50. 7242-7245 (1990)
[文献書誌] Hori,Y.: "Activation of the serumーresponse and TPAーresponse elements by expression of the vーabl protein:comparison of the mode of action of the vーabl protein with those of protein kinase C,cyclic AMPーdependent protein kinase,and the activated cーraf protein." Oncogene. 5. 1201-1206 (1990)
[文献書誌] Fukumoto,Y.: "Molecular cloning and characterization of a novel type of regulatory protein (GDI) for the rho proteins,ras p21ーlike small GTPーbinding proteins." Oncogene. 5. 1321-1328 (1990)
[文献書誌] Matsui,Y.: "Molecular cloning of smg p21B and identification of smg p21 purified from bovine brain and human platelets as smg p21B." Biochem.Biophys.Res.Commun.166. 1010-1016 (1990)
[文献書誌] Fujioka,H.: "A small GTPーbinding protein (G protein) recognized by smg p25A GDP dissociation inhibitor (GDI) in human platelet membranes and GDI for this small G protein in human platelet cytosol." Biochem.Biophys.Res.Commun.168. 1244-1252 (1990)
[文献書誌] Isomura,M.: "Partial purification and characterization of GDP dissociation stimulator (GDS) for the rho proteins from bovine brain cytosol." Biochem.Biophys.Res.Commun.169. 652-659 (1990)
[文献書誌] Ohmori,T.: "Purification and characterization of a novel GTPーbinding protein with a M_r value of 24,000 from rat liver." Biochem.Biophys.Res.Commun.169. 816-823 (1990)
[文献書誌] Kawahara,Y.: "Identification of a major GTPーbinding protein in bovine aortic smooth muscle cytosol as the rhoA gene product." Biochem.Biophys.Res.Commun.170. 673-683 (1990)
[文献書誌] Oku,N.: "Comparison of the mode of action of ras p21 with those of protein kinases A and C in the stimulation of gene expression in NIH/3T3 cells." FEBS.Lett.269. 148-152 (1990)
[文献書誌] Hiroyoshi,M.: "Role or the Cーterminal region of smg p21,a ras p21ーlike small GTPーbinding protein,in membrane and smg p21 GDP/GTP exchange protein interactions." J.Biol.Chem.266. 2962-2969 (1991)
[文献書誌] Hata,Y.: "Enhancement of the actions of smg p21 GDPーGTP exchange protein by the protein kinase Aーcatalyzed phosphorylation of smg p21." J.Biol.Chem.(1991)
[文献書誌] Katayama,M.: "The postーtranslationally modified Cーterminal structure of bovine aortic smooth muscle rhoA p21." J.Biol.Chem.(1991)
[文献書誌] Araki,S.: "Role of the Cーterminal region of smg p25A in its interaction with membranes and the GDP/GTP exchange protein." Mol.Cell.Biol.(1991)
[文献書誌] Sasaki,T.: "A mammalian inhibitory GDP/GTP exchange protein (GDI) for smg p25A is active on the yeast SEC4 protein." Mol.Cell.Biol.(1991)
[文献書誌] Kaibuchi,K.: "Molecular cloning and characterization of the stimulatory GDP/GTP exchange protein (GDS) for smg p21s,ras p21ーlike small GTPーbinding proteins." Mol.Cell.Biol.(1991)
[文献書誌] Isomura,M.: "Regulation of binding of rhoB p20 to membranes by its specific regulatory protein,GDP dissociation inhibitor." Oncogene. (1991)
[文献書誌] Hori,Y.: "Postーtranslational modifications of the Cーterminal region of the rho protein are important for its interaction with membranes and the stimulatory and inhibitory GDP/GTP exchange proteins." Oncogene. (1991)
[文献書誌] Kawata,M.: "The molecular heterogenity of the smgー21/Krevー1/rap1 proteins,a GTPーbinding protein having the same effector domain as ras p21s,in bovine aortic smooth muscle membranes." Oncogene. (1991)
[文献書誌] Ueda,T.: "Purification and characterization from rat liver cytosol of a GDP dissociation inhibitor (GDI) for liver 24KG,a ras p21ーlike GTPーbinding protein,with properties similar to those of smg p25A GDI." Biochemistry. (1991)
[文献書誌] Motoike,T.: "Expression of smg p25A,a ras p21ーlike small GTPーbinding protein,during postnatal development of rat cerebellum." Dev.Brain Res. (1991)
[文献書誌] Kswamura,S.: "Stoichiometric interaction of smg p21 with its GDP/DTP exchange protein and its novel action to regulate the translocation of smg p21 between membrane and cytoplasm." Biochem.Biophys.Commun.(1991)
[文献書誌] Shimizu,K.: "Tissue and subcellular distributions of an inhibitory GDP/GTP exchange protein (GDI) for the rho protein by use of its specific antibody." Biochem.Biophys.Commun.(1991)
[文献書誌] Yoshida,Y.: "A geranylgeranyltransferase for rhoA p21 distinct from the farnesylーtransferase for ras p21s." Biochem.Biophys.Res.Commun.(1991)

1990 年度 実績報告書

低分子量GTP結合蛋白質を介する新しい細胞内情報伝達機構

研究代表者

高井 義美 神戸大学, 医学部, 教授 (60093514)

研究成果

[文献書誌] Fukumoto,Y.: "Activation of the cーfos serumーresponse element by the activated cーHaーras protein in a manner independent of protein kinase C and cAMPーdependent protein kinase." J.Biol.Chem.265. 774-780 (1990)

[文献書誌] Sasaki,T.: "Purification and characterization from bovine cytosol of a protein that inhibits the dissociation of GDP from and the subsequent binding of GTP to smg p25A,a ras p21ーlike GTPーbinding protein." J.Biol.Chem.265. 2333-2337 (1990)

[文献書誌] Hata,Y.: "Inhibition of the ras p21 GTPaseーactivating proteinーstimulated GTPase activity of cーHaーras p21 by smg p21 having the same putative effector domain as ras p21s." J.Biol.Chem.265. 7104-7107 (1990)

[文献書誌] Ueda,T.: "Purification and characterization from bovine brain cytosol of a novel regulatory protein inhibiting the dissociation of GDP from and the subsequent binding of GTP to rhoB p20,a ras p21ーlike GTPーbinding protein." J.Biol.Chem.265. 9373-9380 (1990)

[文献書誌] Mizoguchi,A.: "Localization and subcellular distribution of smg p25A,a ras p21ーlike GTPーbinding protein,in rat brain." J.Biol.Chem.265. 11872-11879 (1990)

[文献書誌] Araki,S.: "Regulation of reversible binding of smg p25A,a ras p21ーlike GTPーbinding protein,to synaptic plasma membranes and vesicles by its regulatory protein,GDP dissociation inhibitor." J.Biol.Chem.265. 13007-13015 (1990)

[文献書誌] Yamamoto,T.: "Purification and characterization from bovine brain cytosol of proteins that regulate the GDP/GTP exchange reaction of smg p21s,ras p21ーlike GTPーbinding proteins." J.Biol.Chem.265. 16626-16634 (1990)

[文献書誌] Kim,S.: "Tissue and subcellular distributions of the smg 21/rap1/Krevー1 proteins which are partly distinct from those of cーras p21s" Mol.Cell.Biol.10. 2645-2652 (1990)

[文献書誌] Matsui,Y.: "Molecular cloning and characterization of a novel type of regulatory protein (GDI) for smg p25A,a ras p21ーlike GTPーbinding protein." Mol.Cell.Biol.10. 4116-4122 (1990)

[文献書誌] Hoshijima,M.: "Purfication and characterization from bovine brain membranes of a GTPーbinding protein with a Mr of 21,000 ADPーribosylated by an ADPーribosyltransferase contaminated in botulinum toxin type C1ーidentification as the rhoA gene product." Mol.Brain Res.7. 9-16 (1990)

[文献書誌] Yamamoto,J.: "A GtPaseーactivating protein for rhoB p20,a ras p21ーlike GTPーbinding proteinーpartial purification,characterization and subcellular distribution in rat brain." Mol.Brain Res.8. 105-111 (1990)

[文献書誌] Sano,K.: "Differential expreession of the Nーmyc,cーfos,and smg p25A genes in human neuroblastoma cells during neuronal and Schwannian differentiation." Mol.Brain Res.9. 149-152 (1990)

[文献書誌] Kawata,M.: "Posttranslationally processed structure of the human platelet protein smg p21B:Evidence for geranylgeranylation and carboxyl methylation of the Cーterminal cysteine." Proc.Natl.Acad.Sci.USA. 87. 8960-8964 (1990)

[文献書誌] Sano,K.: "Expression of the smg p25A (a ras p21ーlike GTPーbinding protein) gene in human neuroblastoma cell lines and tumor tissues." Canser Res.50. 7242-7245 (1990)

[文献書誌] Fukumoto,Y.: "Molecular cloning and characterization of a novel type of regulatory protein (GDI) for the rho proteins,ras p21ーlike small GTPーbinding proteins." Oncogene. 5. 1321-1328 (1990)

[文献書誌] Matsui,Y.: "Molecular cloning of smg p21B and identification of smg p21 purified from bovine brain and human platelets as smg p21B." Biochem.Biophys.Res.Commun.166. 1010-1016 (1990)

[文献書誌] Fujioka,H.: "A small GTPーbinding protein (G protein) recognized by smg p25A GDP dissociation inhibitor (GDI) in human platelet membranes and GDI for this small G protein in human platelet cytosol." Biochem.Biophys.Res.Commun.168. 1244-1252 (1990)

[文献書誌] Isomura,M.: "Partial purification and characterization of GDP dissociation stimulator (GDS) for the rho proteins from bovine brain cytosol." Biochem.Biophys.Res.Commun.169. 652-659 (1990)

[文献書誌] Ohmori,T.: "Purification and characterization of a novel GTPーbinding protein with a M_r value of 24,000 from rat liver." Biochem.Biophys.Res.Commun.169. 816-823 (1990)

[文献書誌] Kawahara,Y.: "Identification of a major GTPーbinding protein in bovine aortic smooth muscle cytosol as the rhoA gene product." Biochem.Biophys.Res.Commun.170. 673-683 (1990)

[文献書誌] Oku,N.: "Comparison of the mode of action of ras p21 with those of protein kinases A and C in the stimulation of gene expression in NIH/3T3 cells." FEBS.Lett.269. 148-152 (1990)

[文献書誌] Hiroyoshi,M.: "Role or the Cーterminal region of smg p21,a ras p21ーlike small GTPーbinding protein,in membrane and smg p21 GDP/GTP exchange protein interactions." J.Biol.Chem.266. 2962-2969 (1991)

[文献書誌] Hata,Y.: "Enhancement of the actions of smg p21 GDPーGTP exchange protein by the protein kinase Aーcatalyzed phosphorylation of smg p21." J.Biol.Chem.(1991)

[文献書誌] Katayama,M.: "The postーtranslationally modified Cーterminal structure of bovine aortic smooth muscle rhoA p21." J.Biol.Chem.(1991)

[文献書誌] Araki,S.: "Role of the Cーterminal region of smg p25A in its interaction with membranes and the GDP/GTP exchange protein." Mol.Cell.Biol.(1991)

[文献書誌] Sasaki,T.: "A mammalian inhibitory GDP/GTP exchange protein (GDI) for smg p25A is active on the yeast SEC4 protein." Mol.Cell.Biol.(1991)

[文献書誌] Kaibuchi,K.: "Molecular cloning and characterization of the stimulatory GDP/GTP exchange protein (GDS) for smg p21s,ras p21ーlike small GTPーbinding proteins." Mol.Cell.Biol.(1991)

[文献書誌] Isomura,M.: "Regulation of binding of rhoB p20 to membranes by its specific regulatory protein,GDP dissociation inhibitor." Oncogene. (1991)

[文献書誌] Hori,Y.: "Postーtranslational modifications of the Cーterminal region of the rho protein are important for its interaction with membranes and the stimulatory and inhibitory GDP/GTP exchange proteins." Oncogene. (1991)

[文献書誌] Kawata,M.: "The molecular heterogenity of the smgー21/Krevー1/rap1 proteins,a GTPーbinding protein having the same effector domain as ras p21s,in bovine aortic smooth muscle membranes." Oncogene. (1991)

[文献書誌] Ueda,T.: "Purification and characterization from rat liver cytosol of a GDP dissociation inhibitor (GDI) for liver 24KG,a ras p21ーlike GTPーbinding protein,with properties similar to those of smg p25A GDI." Biochemistry. (1991)

[文献書誌] Motoike,T.: "Expression of smg p25A,a ras p21ーlike small GTPーbinding protein,during postnatal development of rat cerebellum." Dev.Brain Res. (1991)

[文献書誌] Kswamura,S.: "Stoichiometric interaction of smg p21 with its GDP/DTP exchange protein and its novel action to regulate the translocation of smg p21 between membrane and cytoplasm." Biochem.Biophys.Commun.(1991)

[文献書誌] Shimizu,K.: "Tissue and subcellular distributions of an inhibitory GDP/GTP exchange protein (GDI) for the rho protein by use of its specific antibody." Biochem.Biophys.Commun.(1991)

[文献書誌] Yoshida,Y.: "A geranylgeranyltransferase for rhoA p21 distinct from the farnesylーtransferase for ras p21s." Biochem.Biophys.Res.Commun.(1991)

1990 年度実績報告書

高井義美神戸大学, 医学部, 教授 (60093514)