1991 年度実績報告書

低分子量GTP結合蛋白質の機能修飾物質の開発と有効利用

研究課題

研究課題/領域番号	03558019
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	佐野公彦神戸大学, 医学部, 助手 (40205993) 川原康洋神戸大学, 医学部附属病院, 助手 (80169755) 河田正仁神戸大学, 医学部, 助手 (20224785) 菊池章神戸大学, 医学部, 助手 (10204827) 貝淵弘三神戸大学, 医学部, 助教授 (00169377)
キーワード	低分子量GTP結合蛋白質 / プレニル化 / 翻訳後修飾 / smg GDS / C末端側ペプチド / ファルネシル基転移酵素 / ゲラニルゲラニル基転移酵素
研究概要	私共はすでにsmg p21のGDP/GTP交換反応調節蛋白質(smg GDS)のcDNAをクロ-ニングし、その一次構造を決定している。本年度、私共はsmg GDSがsmg p21のみならずc-ki-ras p21やrhoA p21にも作用することを明らかにした。また、smg GDSがc-ki-ras p21と協同して細胞のトランスフォ-メ-ションを引き起こすことも明らかにした。これらの低分子量G蛋白質はそのC末端側のシステイン残基がプレニル化され翻訳後修飾をうけることが、私共の教室を含めたいくつかの研究室から報告されている。私共はこのC末端の翻訳後修飾がsmg GDSの作用に必須であることを見出した。そこで、smg p21のC末端側のアミノ酸配列をもとにプレニル化されたC末端側ペプチドを合成してsmg GDSに対する作用を検討した。この合成ペプチドはsmg GDSと1対1の比率で結合してsmg GDSの作用を阻害した。今後はこのC末端側ペプチドを用いて細胞や個体レベルでのsmg GDSやsmg p21、c-ki-ras p21、rho p21の生理機能を解析していく予定である。一方、ras p21のC末端領域の翻訳後修飾がras p21のトランスフォ-メ-ション活性の発現に必須であることが報告されている。私共は低分子量G蛋白質の無細胞系でのプレニル化のアッセイ系を確立した。さらに、ras p21のファルネシル基転移酵素、smg p21/rhoA p21のゲラニルゲラニル基転移酵素、smg p25Aのゲラニルゲラニル基転移酵素を分離・精製した。今後はこの酵素群の活性を制御する物質を検索しこれらの低分子量G蛋白質の機能と作用機構の解明に有効利用する予定である。

研究成果

(18件)

すべてその他

すべて文献書誌 (18件)

[文献書誌] Hiroyoshi,M.: "Role of the C-terminal region of smg p21,a ras p21-like small GTP-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/GTP exchange protein by the protein kinase A-catalyzed phosphorylation of smg p21." J.Biol.Chem.266. 6571-6577 (1991)
[文献書誌] Katayama,M.: "The posttranslationally modified C-terminal structure of bovine aortic smooth muscle rhoA p21." J.Biol.Chem.266. 12639-12645 (1991)
[文献書誌] Horiuchi,H.: "A novel prenyltransferase for a small GTP-binding protein having a C-terminal Cys-Ala-Cys structure." J.Biol.Chem.266. 16981-16984 (1991)
[文献書誌] 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-terminal region." J.Biol.Chem.266. 20672-20677 (1991)
[文献書誌] Araki,S.: "Role of the C-terminal region of smg p25A in its interaction with membranes and the GDP/GTP exchange protein." Mol.Cell.Biol.11. 1438-1447 (1991)
[文献書誌] Kaibuchi,K.: "Molecular cloning of the cDNA for stimulatory GDP/GTP exchange protein for smg p21s (ras p21-like small GTP-binding proteins)and characterization of stimulatory GDP/GTP exchange protein." Mol.Cell.Biol.11. 2873-2880 (1991)
[文献書誌] 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)
[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-like GTP-binding protein,with properties similar to those of smg p25A GDI." Biochemistry. 30. 909-917 (1991)
[文献書誌] Farnsworth,C.C.: "C terminus of the small GTP-binding protein smg p25A contains two geranylgeranylated cysteine residues and a methyl ester." Proc.Natl.Acod.Sci.USA. 88. 6196-6200 (1991)
[文献書誌] Mizuno,T.: "A stimulatory GDP/GTP exchange protein for smg p21 is active on the post-translationally processed form of c-Ki-ras p21 and rhoA p21." Proc.Natl.Acod.Sci.USA. 88. 6442-6446 (1991)
[文献書誌] Isomura,M.: "Regulation of binding of rhoB p20 to membranes by its specific regulatory protein,GDP dissociation inhibitor." Oncogene. 6. 119-124 (1991)
[文献書誌] Hori,Y.: "Post-translational modifications of the C-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)
[文献書誌] Kawata,M.: "The molecular heterogeneity of the smg-21/Krev-1/rap1 proteins,a GTP-binding protein having the same effector domain as ras p21s,in bovine aortic smooth muscle membranes." Oncogene. 6. 841-848 (1991)
[文献書誌] Sano,K.: "Differential expression of the N-myc,c-fos,and smg p25A genes in human neuroblastoma cells during neuronal and Schwannian differentiation." Mol.Brain Res.9. 149-152 (1991)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] Fujioka,H.: "Transforming and c-fos promoter/enhancer-stimulating activities of a stimulatory GDP/GTP exchange protein for small GTP-binding proteins." J.Biol.Chem.267. 926-930 (1992)

1991 年度 実績報告書

低分子量GTP結合蛋白質の機能修飾物質の開発と有効利用

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] Hiroyoshi,M.: "Role of the C-terminal region of smg p21,a ras p21-like small GTP-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/GTP exchange protein by the protein kinase A-catalyzed phosphorylation of smg p21." J.Biol.Chem.266. 6571-6577 (1991)

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

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

[文献書誌] 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-terminal region." J.Biol.Chem.266. 20672-20677 (1991)

[文献書誌] Araki,S.: "Role of the C-terminal region of smg p25A in its interaction with membranes and the GDP/GTP exchange protein." Mol.Cell.Biol.11. 1438-1447 (1991)

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

[文献書誌] 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)

[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-like GTP-binding protein,with properties similar to those of smg p25A GDI." Biochemistry. 30. 909-917 (1991)

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

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

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

[文献書誌] Hori,Y.: "Post-translational modifications of the C-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)

[文献書誌] Kawata,M.: "The molecular heterogeneity of the smg-21/Krev-1/rap1 proteins,a GTP-binding protein having the same effector domain as ras p21s,in bovine aortic smooth muscle membranes." Oncogene. 6. 841-848 (1991)

[文献書誌] Sano,K.: "Differential expression of the N-myc,c-fos,and smg p25A genes in human neuroblastoma cells during neuronal and Schwannian differentiation." Mol.Brain Res.9. 149-152 (1991)

[文献書誌] 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)

[文献書誌] 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)

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

1991 年度実績報告書

高井義美神戸大学, 医学部, 教授 (60093514)