1992 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, 医学部附属病院, 講師 (40205993) 川原康洋神戸大学, 医学部附属病院, 講師 (80169755) 佐々木卓也神戸大学, 医学部, 助手 (40241278) 菊池章神戸大学, 医学部, 講師 (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 / 翻訳後修飾 / rho p21 / rho GDI / ras p21 / REKS / rac p21
研究概要	私共はすでに種々の低分子量GTP結合蛋白質(G蛋白質)とその活性制御蛋白質を見出している。低分子量G蛋白質はそのC末端のシステイン残基が脂質による翻訳後修飾を受けることが知られている。また、この翻訳後修飾はG蛋白質が細胞膜に結合する際に必須であることも知られている。私共はすでに、低分子量G蛋白質に活性制御蛋白質が作用する際に、この翻訳後修飾が必須であると明らかにしている。本年度、私共は低分子量G蛋白質の生理機能に対するこの翻訳後修飾の意義を解析した。私共はまずrhop21の活性化を阻害するrho GDIを種々の細胞へマイクロインジェクションすることにより、rho p21が細胞形態の維持、細胞運動、細胞質分裂を制御していることを明らかにした。rho p21がこれらの機能を発現するためにはこの翻訳後修飾が必須であった。さらに、私共はツメガエル卵よりras p21に依存してMAPキナーゼを活性化する因子を部分精製してREKSと命名したが、ras p21がREKSを活性化する際にもras p21の翻訳後修飾が必要であった。また、ras p21は発芽酵母のアデニル酸シクラーゼを活性化するが、この際にもras p21の翻訳後修飾が必須であった。一方、私共はrac p21が好中球のNADPHオキシダーゼを活性化して活性酵素の産生を制御していることを見出しているが、この際にもrac p21の翻訳後修飾が必要であった。したがって、本年度の本研究により、低分子量G蛋白質が生理機能を発現する際にも翻訳後修飾が必須であることが明らかになった。よって、翻訳後修飾に関与する酵素群を制御する物質を開発することによって、低分子量G、蛋白質の生理活性を制御できる可能性が高い。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。

研究成果
(30件)

すべてその他

すべて文献書誌 (30件)

[文献書誌] Miura,Y.: "Regulation of morphology by rho p21 and its inhibitory GDP/GTP exchange protein(rho GDI)in Swiss 3T3 cells." J.Biol.Chem.268. 510-515 (1993)
[文献書誌] Itoh,T.: "The post-translationally processing of ras p21 is critical for its stimulation of Mitogen-activated protein(MAP)kinase." J.Biol.Chem.in press. (1993)
[文献書誌] Takaishi,K.: "Involvement of rho p21 and its inhibitory GDP/GTP exchange protein(rho GDI)in cell motility." Mol.Cell.Biol.13. 72-79 (1993)
[文献書誌] Shirataki,H.: "Rabphilin-3A,a putative target protein for smg p25A/rab3A p25 small GTP-binding protein related to synaptotagmin." Mol.Cell.Biol.in press. (1993)
[文献書誌] Kishi,K.: "Regulation of cytoplasmic division of Xenopus embryo by rho p21 and its inhibitory GDP/GTP exchange protein(rho GDI)." J.Cell Biol.in press. (1993)
[文献書誌] Itoh,T.: "A novel protein factor for ras p21-dependent activation of Mitogen-activated protein kinase." Proc.Natl.Acad.Sci.USA. 90. 975-979 (1993)
[文献書誌] Kawamura,M.: "Translocation of Ki-ras p21 between membrane and cytoplasm by smg GDS." Biochem.Biophys.Res.Commun.in press. (1993)
[文献書誌] Ando,S.: "Post-translational processing of rac p21s is important both for their interaction with the GDP/GTP exchange proteins and for their activation of NADPH oxidase." J.Biol.Chem.267. 25709-25713 (1992)
[文献書誌] Regazzi,R.: "The small GTP-binding proteins in the cytosol of insulin-secreting cells are complexed to GDP dissociation inhibitor proteins." J.Biol.Chem.267. 17512-17519 (1992)
[文献書誌] Kikuchi,A.: "Functional interactions of stimulatory and inhibitory GDP/GTP exchange proteins and their common substrate small GTP-binding protein." J.Biol.Chem.267. 14611-14615 (1992)
[文献書誌] Shirataki,H.: "A possible target protein for smg-25A/rab3A small GTP-binding protein." J.Biol.Chem.267. 10946-10949 (1992)
[文献書誌] Mizuno,T.: "Regulation of the superoxide-generating NADPH oxidase by a small GTP-binding protein and its stimulatory and inhibitory GDP/GTP exchange proteins." J.Biol.Chem.267. 10215-10218 (1992)
[文献書誌] Musha,T.: "The geranylgeranyl moiety but not the methyl moiety of the smg-25A/rab3A protein is essential for the interactions with membrane and its inhibitory GDP/GTP exchange protein." J.Biol.Chem.267. 9821-9825 (1992)
[文献書誌] Hirata,K.: "Involvement of rho p21 in the GTP-enhanced calcium ion sensitivity of smooth muscle contraction." J.Biol.Chem.267. 8719-8722 (1992)
[文献書誌] Sugai,M.: "Epidermal cell differentiation inhibitor ADP-ribosylates small GTP-binding proteins and induces hyperplasia in epidermis." J.Biol.Chem.267. 2600-2604 (1992)
[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)
[文献書誌] Horiuchi,H.: "The posttranslational processing of ras p21 is critical for its stimulation of yeast adenylate cyclase." Mol.Cell.Biol.12. 4515-4520 (1992)
[文献書誌] Yoshida,Y.: "Microinjection of smg/rap1/Krev-1 p21 into Swiss 3T3 cells induces DNA synthesis and morphological changes." Mol.Cell.Biol.12. 3407-3414 (1992)
[文献書誌] Ruggieri,R.: "RSR1,a ras-like gene homologous to Krev-1(smg21A/rap1A):Role in the development of cell polarity and interactions with the Ras pathway in Saccharomyces cerevisiae." Mol.Cell.Biol.12. 758-766 (1992)
[文献書誌] Sakoda,T.: "smg/rap1/Krev-1 p21s inhibit the signal pathway to the c-fos promoter/enhancer from c-Ki-ras p21 but not from c-raf-1 kinase in NIH3T3 cells." Oncogene. 7. 1705-1711 (1992)
[文献書誌] Kotani,K.: "The functional domain of the stimulatory GDP/GTP exchange protein (smg GDS)which interacts with the C-terminal geranylgeranylated region of rap1/Krev-1/smg p21." Oncogene. 7. 1699-1704 (1992)
[文献書誌] Adachi,M.: "Induction of smg p21/rap1A p21/Krev-1 p21 gene expression during phorbol ester-induced differentiation of a human megakaryocytic leukemia cell line." Oncogene. 7. 323-329 (1992)
[文献書誌] Kikuchi,A.: "Molecular cloning of the human cDNA for a stimulatory GDP/GTP exchange protein for c-Ki-ras p21 and smg p21." Oncogene. 7. 289-293 (1992)
[文献書誌] Miura,Y.: "Phosphorylation of smg p21B/rap1B p21 by cyclic GMP-dependent protein kinase." FEBS Lett.297. 171-174 (1992)
[文献書誌] Mizoguchi,A.: "Localization of smg p25A/rab3A p25,a small GTP-binding protein,at the active zone of the rat neuromuscular junction." Biochem.Biophys.Res.Commun.186. 1345-1352 (1992)
[文献書誌] Kuroda,S.: "Cooperative function of rho GDS and rho GDI to regulate rho p21 activation in smooth muscle." Biochem.Biophys.Res.Commun.185. 473-480 (1992)
[文献書誌] Kim,Y.S.: "Effect of chlorpromazine on the smg GDS action." Biochem.Biophys.Res.Commun.182. 1446-1453 (1992)
[文献書誌] Hiraoka,K.: "Both stimulatory and inhibitory GDP/GTP exchange proteins,smg GDS and rho GDI,are active on multiple small GTP-binding proteins." Biochem.Biophys.Res.Commun.182. 921-930 (1992)
[文献書誌] Regazzi,R.: "Characterization of small-molecular-mass guanine-nucleotide-binding regulatory proteins in insulin-secreting cells and PC12 cells." Eur.J.Biochem.208. 729-737 (1992)
[文献書誌] Takai,Y.: "Small GTP-binding proteins." Int.Rev.Cytol.133. 187-230 (1992)

1992 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] Miura,Y.: "Regulation of morphology by rho p21 and its inhibitory GDP/GTP exchange protein(rho GDI)in Swiss 3T3 cells." J.Biol.Chem.268. 510-515 (1993)

[文献書誌] Itoh,T.: "The post-translationally processing of ras p21 is critical for its stimulation of Mitogen-activated protein(MAP)kinase." J.Biol.Chem.in press. (1993)

[文献書誌] Takaishi,K.: "Involvement of rho p21 and its inhibitory GDP/GTP exchange protein(rho GDI)in cell motility." Mol.Cell.Biol.13. 72-79 (1993)

[文献書誌] Shirataki,H.: "Rabphilin-3A,a putative target protein for smg p25A/rab3A p25 small GTP-binding protein related to synaptotagmin." Mol.Cell.Biol.in press. (1993)

[文献書誌] Kishi,K.: "Regulation of cytoplasmic division of Xenopus embryo by rho p21 and its inhibitory GDP/GTP exchange protein(rho GDI)." J.Cell Biol.in press. (1993)

[文献書誌] Itoh,T.: "A novel protein factor for ras p21-dependent activation of Mitogen-activated protein kinase." Proc.Natl.Acad.Sci.USA. 90. 975-979 (1993)

[文献書誌] Kawamura,M.: "Translocation of Ki-ras p21 between membrane and cytoplasm by smg GDS." Biochem.Biophys.Res.Commun.in press. (1993)

[文献書誌] Ando,S.: "Post-translational processing of rac p21s is important both for their interaction with the GDP/GTP exchange proteins and for their activation of NADPH oxidase." J.Biol.Chem.267. 25709-25713 (1992)

[文献書誌] Regazzi,R.: "The small GTP-binding proteins in the cytosol of insulin-secreting cells are complexed to GDP dissociation inhibitor proteins." J.Biol.Chem.267. 17512-17519 (1992)

[文献書誌] Kikuchi,A.: "Functional interactions of stimulatory and inhibitory GDP/GTP exchange proteins and their common substrate small GTP-binding protein." J.Biol.Chem.267. 14611-14615 (1992)

[文献書誌] Shirataki,H.: "A possible target protein for smg-25A/rab3A small GTP-binding protein." J.Biol.Chem.267. 10946-10949 (1992)

[文献書誌] Mizuno,T.: "Regulation of the superoxide-generating NADPH oxidase by a small GTP-binding protein and its stimulatory and inhibitory GDP/GTP exchange proteins." J.Biol.Chem.267. 10215-10218 (1992)

[文献書誌] Musha,T.: "The geranylgeranyl moiety but not the methyl moiety of the smg-25A/rab3A protein is essential for the interactions with membrane and its inhibitory GDP/GTP exchange protein." J.Biol.Chem.267. 9821-9825 (1992)

[文献書誌] Hirata,K.: "Involvement of rho p21 in the GTP-enhanced calcium ion sensitivity of smooth muscle contraction." J.Biol.Chem.267. 8719-8722 (1992)

[文献書誌] Sugai,M.: "Epidermal cell differentiation inhibitor ADP-ribosylates small GTP-binding proteins and induces hyperplasia in epidermis." J.Biol.Chem.267. 2600-2604 (1992)

[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)

[文献書誌] Horiuchi,H.: "The posttranslational processing of ras p21 is critical for its stimulation of yeast adenylate cyclase." Mol.Cell.Biol.12. 4515-4520 (1992)

[文献書誌] Yoshida,Y.: "Microinjection of smg/rap1/Krev-1 p21 into Swiss 3T3 cells induces DNA synthesis and morphological changes." Mol.Cell.Biol.12. 3407-3414 (1992)

[文献書誌] Ruggieri,R.: "RSR1,a ras-like gene homologous to Krev-1(smg21A/rap1A):Role in the development of cell polarity and interactions with the Ras pathway in Saccharomyces cerevisiae." Mol.Cell.Biol.12. 758-766 (1992)

[文献書誌] Sakoda,T.: "smg/rap1/Krev-1 p21s inhibit the signal pathway to the c-fos promoter/enhancer from c-Ki-ras p21 but not from c-raf-1 kinase in NIH3T3 cells." Oncogene. 7. 1705-1711 (1992)

[文献書誌] Kotani,K.: "The functional domain of the stimulatory GDP/GTP exchange protein (smg GDS)which interacts with the C-terminal geranylgeranylated region of rap1/Krev-1/smg p21." Oncogene. 7. 1699-1704 (1992)

[文献書誌] Adachi,M.: "Induction of smg p21/rap1A p21/Krev-1 p21 gene expression during phorbol ester-induced differentiation of a human megakaryocytic leukemia cell line." Oncogene. 7. 323-329 (1992)

[文献書誌] Kikuchi,A.: "Molecular cloning of the human cDNA for a stimulatory GDP/GTP exchange protein for c-Ki-ras p21 and smg p21." Oncogene. 7. 289-293 (1992)

[文献書誌] Miura,Y.: "Phosphorylation of smg p21B/rap1B p21 by cyclic GMP-dependent protein kinase." FEBS Lett.297. 171-174 (1992)

[文献書誌] Mizoguchi,A.: "Localization of smg p25A/rab3A p25,a small GTP-binding protein,at the active zone of the rat neuromuscular junction." Biochem.Biophys.Res.Commun.186. 1345-1352 (1992)

[文献書誌] Kuroda,S.: "Cooperative function of rho GDS and rho GDI to regulate rho p21 activation in smooth muscle." Biochem.Biophys.Res.Commun.185. 473-480 (1992)

[文献書誌] Kim,Y.S.: "Effect of chlorpromazine on the smg GDS action." Biochem.Biophys.Res.Commun.182. 1446-1453 (1992)

[文献書誌] Hiraoka,K.: "Both stimulatory and inhibitory GDP/GTP exchange proteins,smg GDS and rho GDI,are active on multiple small GTP-binding proteins." Biochem.Biophys.Res.Commun.182. 921-930 (1992)

[文献書誌] Regazzi,R.: "Characterization of small-molecular-mass guanine-nucleotide-binding regulatory proteins in insulin-secreting cells and PC12 cells." Eur.J.Biochem.208. 729-737 (1992)

[文献書誌] Takai,Y.: "Small GTP-binding proteins." Int.Rev.Cytol.133. 187-230 (1992)

1992 年度実績報告書

高井義美神戸大学, 医学部, 教授 (60093514)