1991 Fiscal Year Annual Research Report

低分子量GTP結合蛋白質を介する細胞装情報伝達機構とその病態

Research Project

Project/Area Number	03404022
Research Institution	Kobe University
Principal Investigator	高井義美神戸大学, 医学部, 教授 (60093514)
Co-Investigator(Kenkyū-buntansha)	河田正仁神戸大学, 医学部, 助手 (20224785) 菊地章神戸大学, 医学部, 助手 (10204827) 貝淵弘三神戸大学, 医学部, 助教授 (00169377)
Keywords	低分子量GTP結合蛋白質 / 細胞内情報伝達機構 / 翻訳後修飾 / ゲラニルゲラニル基 / リン酸化 / 平滑筋 / 活性酸素
Research Abstract	昨年度までに私共は.多数の低分子量GTP結合蛋白質(G蛋白質)とその活性制御蛋白質(GDS,GDI,GAP)を見出している。GDSとGDIは低分子量G蛋白質のGDP/GTP交換反応を、GAPはそのGTPase活性を制御する。 ras p21ではそのC末端側の翻訳後修飾がras p21の機能に重要であることが明らかになっているので、本年度は、まず低分子量G蛋白質の翻訳後修飾を解析した。smg p21とrhoA p21のC末端側のアミノ酸配列はCysーAーAーX(Aは脂肪族アミノ酸;Xは任意のアミノ酸)であるが、Cys残基にゲラニルゲラニル基が結合し、続いてC末端側の3個のアミノ酸が除去され最後にCys残基がメチル化されることが明らかとなった。smg p25AのC末端側のアミノ酸配列はCysーXーCysであるが、この両者のCys残基にゲラニルゲラニル基が結合し、C末端側のCys残基はメチル化されていた。これらの低分子量G蛋白質の翻訳後修飾は低分子量G蛋白質の細胞膜への結合とGDSおよびGDIの作用に必須であった。したがって、ras p21以外の低分子量G蛋白質においても.翻訳後修飾が低分子量G蛋白質の機能に重要であることが示唆された。次に.私共は既知の細胞内情報伝達系と低分子量G蛋白質との関連を解析した。smg p21はAキナ-ゼとGキナ-ゼによりリン酸化され、smg p21はリン酸化されるとはじめてsmg GDSにより活性化されることが明らかになった。一方.rho p21は平滑筋収縮におけるCa^<2+>感受性を庄進し、rho p21のこの作用はrho p21をADPリボシル化すると消失した。他方、好中球における活性酸素の産生はCキナ-ゼ系とCa^<2+>系により調節されているが、rog p21はこの活性酸素の産生を促進した。したがって、低分子量G蛋白質は既知の細胞内情報伝達系と密接に関連しながら種々の細胞機能を制御することが明らかになった。以上の結果を得たことにより、本年度の研究計画はほぼ達成できたと考えている。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] Hiroyoshi,M.: "Role of 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)
[Publications] Hata,Y.: "Enhancement of the actions of smg p21 GDP/GTP exchange protein by the protein kinase Aーcatalyzed phosphorylation of smg p21." J.Biol.Chem. 266. 6571-6577 (1991)
[Publications] Katayama,M.: "The posttranslationally modified Cーterminal structure of bovine aortic smooth muscle rhoA p21." J.Biol.Chem.266. 12639-12645 (1991)
[Publications] Horiuchi,H.: "A novel prenyltransferase for a small GTPーbinding protein having a Cーterminal CysーAlaーCys structure." J.Biol.Chem.266. 16981-16984 (1991)
[Publications] Shirataki,H.: "Inhibition of the action of the stimulatory GDP/GTP exchange protein for smg p21 by the geranylgeranylated synthetic peptides designed from its Cーterminal region." J.Biol.Chem.266. 20672-20677 (1991)
[Publications] 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.11. 1438-1447 (1991)
[Publications] Kaibuchi,K.: "Molecular cloning of the cDNA for stimulatory GDP/GTP exchange protein for smg p21s(ras p21ーlike small GTPーbinding proteins)and characterization of stimulatory GDP/GTP exchange protein." Mol.Cell.Biol.11. 2873-2880 (1991)
[Publications] Sasaki,T.: "A mammalian inhibitory GDP/GTP exchange protein(GDP dissociation inhibitor)for smg p25A is active on the yeast SEC4 protein." Mol.Cell.Biol.11. 2909-2912 (1991)
[Publications] 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. 30. 909-917 (1991)
[Publications] Farnsworth,C.C.: "C terminus of the small GTPーbinding protein smg p25A contains two geranylgeranylated cysteine residues and a methyl ester." Proc.Natl.Acod.Sci.USA. 88. 6196-6200 (1991)
[Publications] Mizuno,T.: "A stimulatory GDP/GTP exchange protein for smg p21 is active on the postーtranslationally processed form of cーKiーras p21 and rhoA p21." Proc.Natl.Acod.Sci.USA. 88. 6442-6446 (1991)
[Publications] Isomura,M.: "Regulation of binding of rhoB p20 to membranes by its specific regulatory protein,GDP dissociation inhibitor." Oncogene. 6. 119-124 (1991)
[Publications] 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. 6. 515-522 (1991)
[Publications] Kawata,M.: "The molecular heterogeneity 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. 6. 841-848 (1991)
[Publications] Sano,K.: "Differential expression 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 (1991)
[Publications] Takayama,K.: "Similarity of expression of low molecular weight G proteins smg p21A and ras p21 in normal and malignant human tissues." Cancer Res.51. 2223-2228 (1991)
[Publications] Kawamura,S.: "Inhibition of the action of a stimulatory GDP/GTP exchange protein for smg p21 by acidic membrane phospholipids." Jpn.J.Cancer Res.82. 758-761 (1991)
[Publications] Fujioka,H.: "Transforming and Cーfos promoter/enhancerーstimulating activities of a stimulatory GDP/GTP exchange protein for small GTPーbinding proteins." J.Biol.Chem.267. 926-930 (1992)

1991 Fiscal Year Annual Research Report

低分子量GTP結合蛋白質を介する細胞装情報伝達機構とその病態

Principal Investigator

高井 義美 神戸大学, 医学部, 教授 (60093514)

Research Products

[Publications] Hiroyoshi,M.: "Role of 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)

[Publications] Hata,Y.: "Enhancement of the actions of smg p21 GDP/GTP exchange protein by the protein kinase Aーcatalyzed phosphorylation of smg p21." J.Biol.Chem. 266. 6571-6577 (1991)

[Publications] Katayama,M.: "The posttranslationally modified Cーterminal structure of bovine aortic smooth muscle rhoA p21." J.Biol.Chem.266. 12639-12645 (1991)

[Publications] Horiuchi,H.: "A novel prenyltransferase for a small GTPーbinding protein having a Cーterminal CysーAlaーCys structure." J.Biol.Chem.266. 16981-16984 (1991)

[Publications] Shirataki,H.: "Inhibition of the action of the stimulatory GDP/GTP exchange protein for smg p21 by the geranylgeranylated synthetic peptides designed from its Cーterminal region." J.Biol.Chem.266. 20672-20677 (1991)

[Publications] 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.11. 1438-1447 (1991)

[Publications] Kaibuchi,K.: "Molecular cloning of the cDNA for stimulatory GDP/GTP exchange protein for smg p21s(ras p21ーlike small GTPーbinding proteins)and characterization of stimulatory GDP/GTP exchange protein." Mol.Cell.Biol.11. 2873-2880 (1991)

[Publications] Sasaki,T.: "A mammalian inhibitory GDP/GTP exchange protein(GDP dissociation inhibitor)for smg p25A is active on the yeast SEC4 protein." Mol.Cell.Biol.11. 2909-2912 (1991)

[Publications] 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. 30. 909-917 (1991)

[Publications] Farnsworth,C.C.: "C terminus of the small GTPーbinding protein smg p25A contains two geranylgeranylated cysteine residues and a methyl ester." Proc.Natl.Acod.Sci.USA. 88. 6196-6200 (1991)

[Publications] Mizuno,T.: "A stimulatory GDP/GTP exchange protein for smg p21 is active on the postーtranslationally processed form of cーKiーras p21 and rhoA p21." Proc.Natl.Acod.Sci.USA. 88. 6442-6446 (1991)

[Publications] Isomura,M.: "Regulation of binding of rhoB p20 to membranes by its specific regulatory protein,GDP dissociation inhibitor." Oncogene. 6. 119-124 (1991)

[Publications] 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. 6. 515-522 (1991)

[Publications] Kawata,M.: "The molecular heterogeneity 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. 6. 841-848 (1991)

[Publications] Sano,K.: "Differential expression 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 (1991)

[Publications] Takayama,K.: "Similarity of expression of low molecular weight G proteins smg p21A and ras p21 in normal and malignant human tissues." Cancer Res.51. 2223-2228 (1991)

[Publications] Kawamura,S.: "Inhibition of the action of a stimulatory GDP/GTP exchange protein for smg p21 by acidic membrane phospholipids." Jpn.J.Cancer Res.82. 758-761 (1991)

[Publications] Fujioka,H.: "Transforming and Cーfos promoter/enhancerーstimulating activities of a stimulatory GDP/GTP exchange protein for small GTPーbinding proteins." J.Biol.Chem.267. 926-930 (1992)

高井義美神戸大学, 医学部, 教授 (60093514)