1997 Fiscal Year Annual Research Report

シグナル伝達系におけるレセプター・G蛋白質相互作用の原子レベルでの解析

Research Project

Project/Area Number	08308038
Research Institution	Kyoto University
Principal Investigator	七田芳則京都大学, 大学院・理学研究科, 助教授 (60127090)
Co-Investigator(Kenkyū-buntansha)	藤吉好則京都大学, 大学院・理学研究科, 教授 (80142298) 徳永史生大阪大学, 理学部, 教授 (80025452) 津田基之姫路工業大学, 理学部, 教授 (60045458) 加茂直樹北海道大学, 薬学部, 教授 (10001976) 垣谷俊昭名古屋大学, 大学院・理学研究科, 教授 (90027350)
Keywords	受容体 / G蛋白質 / ロドプシン / 先 / バクテリオロドプシン / 情報 / 構造 / 機能
Research Abstract	受容体-G蛋白質相互作用に基づく細胞内情報伝達機構について、光情報を伝達するロドプシン-トランスデューシン(Gt)相互作用をモデル系として検討した結果、以下の知見が得られた。 1.ウシロドプシンがG蛋白質と相互作用する動的過程を分光学的に解析した結果、結合と活性化に対応する二つの中間体が存在することを明らかにした。また、タコロドプシンでもG蛋白質を活性化する中間体を同定した。さらに、相互作用したときの構造変化をFTIRで検出し、GtのC末端の残基による変化を同定した。 2.ウシロドプシンの精製/結晶化条件を更に改善して新たな三次元結晶を得ることに成功した。また、天然の状態に近いGtの2次元結晶を作製して膜表面に吸着する姿勢について解析した結果、X線結晶学を用いて予測された結果とは異なることが明らかになった。 3.古細菌の負の走光性の受容体を大腸菌の膜に発現させることに成功し、中間体を同定した。また、バクテリオロドプシンの光中間体の安定性を変性実験と立体構造予測により検討した結果、M中間体では他の中間体と異なった構造になりやすいという結果が得られた。 4.ハトとヒキガエルの脳から脳深部(視床下部)に存在する光受容体をコードするcDNAを単離することに成功した。また、ヒト赤色光受容体と緑色光受容体の色変化に関与するアミノ酸残基のうち、3つの直接的に効くアミノ酸残基の間の協力現象と、4つの間接的に効くアミノ酸残基の役割があることを明らかにした。 5.視物質のリン酸化反応を制御するSモジュリンの変異体を作成してその性質を調べた結果、膜結合性とカルシウムイオン結合に関与するアミノ酸を同定した。また、ROS内信号伝達についてモデルを用いて検討し、通常の拡散を基本にした信号伝達機構では実験結果を説明できないことを示した。

Research Products
(50 results)

All Other

All Publications (50 results)

[Publications] H.Imai: "Single amino acid residue as a functional determinant of rod and cone pigments." Proc.Natl.Acad.Sci.USA. 94. 2322-2326 (1997)
[Publications] T.Nagata: "Water and peptide bockborn structure in the active center of bovine rhodopsin." Biochemistry. 36. 6164-6170 (1997)
[Publications] H.Imai: "Photochemical and biochemical properties of chicken blue-sensitive cone visual pigment." Biochemistry. 36. 12773-12779 (1997)
[Publications] D.Kojima: "A novel Go-mediated phototransduction cascade in scallop visual cells." J.Biol.Chem.272. 22979-22982 (1997)
[Publications] S.Tachibanaki: "Presence of two rhodopsin intermediates responsible for transducin activation." Biochemistry. 36. 14173-14180 (1997)
[Publications] R.Akiyama: "Analysis of excited state dynamics of 13-trans-locked-bacteriorhodopsin." J.Phys.Chem.101. 412-417 (1997)
[Publications] A.Wada: "Retinoids and related compounds. Part 20. Synthesis of 11-Z-8,18-ethanoretinal and conformational study of rhodopsin chromophore." Chem.Soc.Perkin Trans.1. 1773-1777 (1997)
[Publications] S.Tachibanaki: "Identification of a new intermediate state that binds but not activate transducin in the bleaching process of bovine rhodopsin." FEBS lett.(印刷中). (1998)
[Publications] Y.Shichida: "Visual Pigments." J.Cell,Mol.Life Sci.(印刷中).
[Publications] 今井啓雄: "時間分解低温分光法による蛋白質構造変化の解析" 生物物理. 33. 225-226 (1997)
[Publications] N.Goto: "Monte Carlo Simulation on the structure and reaction at metal-electrode interface." J.Phys.Soc.Jpn.66. 1825-1835 (1997)
[Publications] T.Yamato: "Molecular dynamics simulation of the excited-state dynamics of bacteriorhodopsin." Photochem.Photobiol.66. 735-740 (1997)
[Publications] T.Kakitani: "Deuterium substitution effect on the excited state dynamics of rhodopsin." J.Phys.Chem.(印刷中). (1998)
[Publications] 垣谷俊昭: "多くの車輪が光を仕事に変える" 化学. 52. 68-69 (1997)
[Publications] 垣谷俊昭: "いまなぜ電子移動反応か" 化学. 52. 30-32 (1997)
[Publications] T.Kikukawa: "Restricted motion of photoexcited bacteriorhodopsin in purple membrane containing ethanol." Biophys.J.73. 357-366 (1997)
[Publications] K.Shimono: "Functional expression of pharaonis phoborhodopsin in Escherichia coli." FEBS Letters. 420. 54-56 (1997)
[Publications] M.Nakagawa: "Identification of two palmitoyl groups in octopus rhodopsin." Photochem.Photobiol.65. 187-191 (1997)
[Publications] M.Nakagawa: "Light-induced protein conformational changes in the photolysis of octopus rhodopsin." Biophysical J.72. 2320-2328 (1997)
[Publications] S.Nishimura: "Structural dynamics of waters and the peptide backbone around the Schiff base associated with the light-activated process of octopus rhodopsin." Biochemistry. 36. 864-870 (1997)
[Publications] O.Hisatomi: "Arrestins expressed in killifish photoreceptor cells." FEBS Lett.411(1). 12-18 (1997)
[Publications] S.Matsuda: "Functional expression and characterization of frog photoreceptor-specific calcium-binding proteins." Biochem.Biophys.Res.Commun.234(1). 173-177 (1997)
[Publications] O.Hisatomi: "The primary structure and distribution of killifish visual pigments." Vision Res.37(22). 3089-3096 (1997)
[Publications] Y.Nishiwaki: "Three-dimensional reconstitution of cone arrangement on the spherical surface of the retina in the medaka eyes." Zool.Sci.14(10). 795-801 (1997)
[Publications] Y.Imamoto: "Evidence for proton transfer from Glu-46 to the chromophore during the photocycle of photoactive yellow protein." J.Biol.Chem.272. 12905-12908 (1997)
[Publications] K.Mitsuoka: "Examination of the Leaf Scan 45,a line-illuminating micro-densitometer,for its use in electron crystallography." Ultramicroscopy. 68. 109-121 (1997)
[Publications] T.walz: "Three-dimensional structure of aquaporin 1." Nature. 387. 624-627 (1997)
[Publications] Y.Kimura: "Surface of bacteriorhodopsin revealed by high-resolution electron crystallography." 389. 206-211 (1997)
[Publications] H.Hebert: "The 3.0 A projection structure of microsomal glutathione transferaseas determined by electron crystallography of p21212 two-dimensional crystals." J.Mol.Biol.271. 751-758 (1997)
[Publications] Y.Kimura: "High resolution structure of bacteriorhodopsin determined by electron crystallography." Photochem.Photobiol.66. 764-767 (1997)
[Publications] T.Doi: "Characterization of human endotheine ETB receptor and mutants expressed in insect cells." Eur.J.Biochem.248. 139-148 (1997)
[Publications] Y.Zhu: "Details of the arrangement of the outer capsid of rice dwarf phytoreovirus,as visualized by two-dimensional crystallography." J.Virol.71. 8899-8901 (1997)
[Publications] I.Ikeda: "Projection map of the reaction center-light harvesting complex from Rhodopseudomanas viridis at 10 A resolution." FEBS Letters. (印刷中). (1998)
[Publications] T.Okano: "Immunocytochemical identification of pinopsin in pineal grands of chicken and pigeon." Mol.Brain Res.50. 190-196 (1997)
[Publications] Y.Takanaka: "Light-dependent expression of pinopsin gene in chicken pineal gland." J.Neurochem.(印刷中). (1998)
[Publications] T.Yoshikawa: "A deep brain photoreceptive molecule in the toad hypothalamus." FEBS Lett.(印刷中). (1998)
[Publications] Y.Wada: "Identification of toad deep brain photoreceptor molecule." FEBS Lett.(印刷中).
[Publications] A.Etoh: "Light-Induced denaturation of bacteriorhodopsin just above melting point of two-dimensional crystal." J.Phys.Soc.Jpn.66-4. 975-978 (1997)
[Publications] A.Etoh: "Anomalous ultrasonic adsorption of liposome suspensions in the liquid crystal Phase." Jpn.J.Appl.Phys.36-1-5A. 2922-2928 (1997)
[Publications] M.Masuko: "Optimal configuration of pyrene excimer induced by the excimerforming two-probe nucleic acid hybridization method." Nucleic Acids Symposium Series. 37. 83-84 (1997)
[Publications] M.Hatanaka: "Localization and orientation of functional water molecules in bacteriorhodopsin as revealed by polarized Fourier transform infrared spectroscopy." Biophys.J.73-2. 1001-1006 (1997)
[Publications] S.Nishimura: "Transmembrane signaling mediated by water in bovine rhodopsin." Photochem.Photobiol.66-6. 796-801 (1997)
[Publications] T.Okada: "Highly selective separation of rhodopsin from bovine rod outer segment membranes using combination of divalent cation and alkyl (thio) glucoside." Photochem.Photobiol.(印刷中). (1998)
[Publications] 垣谷俊昭: "光・物質・生命と反応(単著)" 丸善, 206 (1998)
[Publications] 津田基之: "広川ニューロサイエンス6「二次メッセンジャーと産生酵素」(竹縄忠臣、野村靖幸編)" 広川書店, 148 (1997)
[Publications] 津田基之: "生物のスーパーセンサー" 共立出版, 186 (1997)
[Publications] 藤吉好則: "細胞工学" 秀潤社, 13 (1997)
[Publications] 市川一寿: "記憶とは何か記憶の分子生物学(分担執筆)" 丸善, 126 (1997)
[Publications] 七田芳則: "生物物理から見た生命像4 視る感覚「光のセンサー;視物質」(分担執筆)" 吉岡書店, 127 (1997)
[Publications] 七田芳則: "視覚、図で学び絵で見るレーザー科学入門(分担執筆)" 丸善(CD-ROM), (1997)

1997 Fiscal Year Annual Research Report

シグナル伝達系におけるレセプター・G蛋白質相互作用の原子レベルでの解析

Principal Investigator

七田 芳則 京都大学, 大学院・理学研究科, 助教授 (60127090)

Research Products

[Publications] H.Imai: "Single amino acid residue as a functional determinant of rod and cone pigments." Proc.Natl.Acad.Sci.USA. 94. 2322-2326 (1997)

[Publications] T.Nagata: "Water and peptide bockborn structure in the active center of bovine rhodopsin." Biochemistry. 36. 6164-6170 (1997)

[Publications] H.Imai: "Photochemical and biochemical properties of chicken blue-sensitive cone visual pigment." Biochemistry. 36. 12773-12779 (1997)

[Publications] D.Kojima: "A novel Go-mediated phototransduction cascade in scallop visual cells." J.Biol.Chem.272. 22979-22982 (1997)

[Publications] S.Tachibanaki: "Presence of two rhodopsin intermediates responsible for transducin activation." Biochemistry. 36. 14173-14180 (1997)

[Publications] R.Akiyama: "Analysis of excited state dynamics of 13-trans-locked-bacteriorhodopsin." J.Phys.Chem.101. 412-417 (1997)

[Publications] A.Wada: "Retinoids and related compounds. Part 20. Synthesis of 11-Z-8,18-ethanoretinal and conformational study of rhodopsin chromophore." Chem.Soc.Perkin Trans.1. 1773-1777 (1997)

[Publications] S.Tachibanaki: "Identification of a new intermediate state that binds but not activate transducin in the bleaching process of bovine rhodopsin." FEBS lett.(印刷中). (1998)

[Publications] Y.Shichida: "Visual Pigments." J.Cell,Mol.Life Sci.(印刷中).

[Publications] 今井啓雄: "時間分解低温分光法による蛋白質構造変化の解析" 生物物理. 33. 225-226 (1997)

[Publications] N.Goto: "Monte Carlo Simulation on the structure and reaction at metal-electrode interface." J.Phys.Soc.Jpn.66. 1825-1835 (1997)

[Publications] T.Yamato: "Molecular dynamics simulation of the excited-state dynamics of bacteriorhodopsin." Photochem.Photobiol.66. 735-740 (1997)

[Publications] T.Kakitani: "Deuterium substitution effect on the excited state dynamics of rhodopsin." J.Phys.Chem.(印刷中). (1998)

[Publications] 垣谷俊昭: "多くの車輪が光を仕事に変える" 化学. 52. 68-69 (1997)

[Publications] 垣谷俊昭: "いまなぜ電子移動反応か" 化学. 52. 30-32 (1997)

[Publications] T.Kikukawa: "Restricted motion of photoexcited bacteriorhodopsin in purple membrane containing ethanol." Biophys.J.73. 357-366 (1997)

[Publications] K.Shimono: "Functional expression of pharaonis phoborhodopsin in Escherichia coli." FEBS Letters. 420. 54-56 (1997)

[Publications] M.Nakagawa: "Identification of two palmitoyl groups in octopus rhodopsin." Photochem.Photobiol.65. 187-191 (1997)

[Publications] M.Nakagawa: "Light-induced protein conformational changes in the photolysis of octopus rhodopsin." Biophysical J.72. 2320-2328 (1997)

[Publications] S.Nishimura: "Structural dynamics of waters and the peptide backbone around the Schiff base associated with the light-activated process of octopus rhodopsin." Biochemistry. 36. 864-870 (1997)

[Publications] O.Hisatomi: "Arrestins expressed in killifish photoreceptor cells." FEBS Lett.411(1). 12-18 (1997)

[Publications] S.Matsuda: "Functional expression and characterization of frog photoreceptor-specific calcium-binding proteins." Biochem.Biophys.Res.Commun.234(1). 173-177 (1997)

[Publications] O.Hisatomi: "The primary structure and distribution of killifish visual pigments." Vision Res.37(22). 3089-3096 (1997)

[Publications] Y.Nishiwaki: "Three-dimensional reconstitution of cone arrangement on the spherical surface of the retina in the medaka eyes." Zool.Sci.14(10). 795-801 (1997)

[Publications] Y.Imamoto: "Evidence for proton transfer from Glu-46 to the chromophore during the photocycle of photoactive yellow protein." J.Biol.Chem.272. 12905-12908 (1997)

[Publications] K.Mitsuoka: "Examination of the Leaf Scan 45,a line-illuminating micro-densitometer,for its use in electron crystallography." Ultramicroscopy. 68. 109-121 (1997)

[Publications] T.walz: "Three-dimensional structure of aquaporin 1." Nature. 387. 624-627 (1997)

[Publications] Y.Kimura: "Surface of bacteriorhodopsin revealed by high-resolution electron crystallography." 389. 206-211 (1997)

[Publications] H.Hebert: "The 3.0 A projection structure of microsomal glutathione transferaseas determined by electron crystallography of p21212 two-dimensional crystals." J.Mol.Biol.271. 751-758 (1997)

[Publications] Y.Kimura: "High resolution structure of bacteriorhodopsin determined by electron crystallography." Photochem.Photobiol.66. 764-767 (1997)

[Publications] T.Doi: "Characterization of human endotheine ETB receptor and mutants expressed in insect cells." Eur.J.Biochem.248. 139-148 (1997)

[Publications] Y.Zhu: "Details of the arrangement of the outer capsid of rice dwarf phytoreovirus,as visualized by two-dimensional crystallography." J.Virol.71. 8899-8901 (1997)

[Publications] I.Ikeda: "Projection map of the reaction center-light harvesting complex from Rhodopseudomanas viridis at 10 A resolution." FEBS Letters. (印刷中). (1998)

[Publications] T.Okano: "Immunocytochemical identification of pinopsin in pineal grands of chicken and pigeon." Mol.Brain Res.50. 190-196 (1997)

[Publications] Y.Takanaka: "Light-dependent expression of pinopsin gene in chicken pineal gland." J.Neurochem.(印刷中). (1998)

[Publications] T.Yoshikawa: "A deep brain photoreceptive molecule in the toad hypothalamus." FEBS Lett.(印刷中). (1998)

[Publications] Y.Wada: "Identification of toad deep brain photoreceptor molecule." FEBS Lett.(印刷中).

[Publications] A.Etoh: "Light-Induced denaturation of bacteriorhodopsin just above melting point of two-dimensional crystal." J.Phys.Soc.Jpn.66-4. 975-978 (1997)

[Publications] A.Etoh: "Anomalous ultrasonic adsorption of liposome suspensions in the liquid crystal Phase." Jpn.J.Appl.Phys.36-1-5A. 2922-2928 (1997)

[Publications] M.Masuko: "Optimal configuration of pyrene excimer induced by the excimerforming two-probe nucleic acid hybridization method." Nucleic Acids Symposium Series. 37. 83-84 (1997)

[Publications] M.Hatanaka: "Localization and orientation of functional water molecules in bacteriorhodopsin as revealed by polarized Fourier transform infrared spectroscopy." Biophys.J.73-2. 1001-1006 (1997)

[Publications] S.Nishimura: "Transmembrane signaling mediated by water in bovine rhodopsin." Photochem.Photobiol.66-6. 796-801 (1997)

[Publications] T.Okada: "Highly selective separation of rhodopsin from bovine rod outer segment membranes using combination of divalent cation and alkyl (thio) glucoside." Photochem.Photobiol.(印刷中). (1998)

[Publications] 垣谷俊昭: "光・物質・生命と反応(単著)" 丸善, 206 (1998)

[Publications] 津田基之: "広川ニューロサイエンス6「二次メッセンジャーと産生酵素」(竹縄忠臣、野村靖幸 編)" 広川書店, 148 (1997)

[Publications] 津田基之: "生物のスーパーセンサー" 共立出版, 186 (1997)

[Publications] 藤吉好則: "細胞工学" 秀潤社, 13 (1997)

[Publications] 市川一寿: "記憶とは何か 記憶の分子生物学(分担執筆)" 丸善, 126 (1997)

[Publications] 七田芳則: "生物物理から見た生命像4 視る感覚「光のセンサー;視物質」(分担執筆)" 吉岡書店, 127 (1997)

[Publications] 七田芳則: "視覚、図で学び絵で見るレーザー科学入門(分担執筆)" 丸善(CD-ROM), (1997)

七田芳則京都大学, 大学院・理学研究科, 助教授 (60127090)

[Publications] 津田基之: "広川ニューロサイエンス6「二次メッセンジャーと産生酵素」(竹縄忠臣、野村靖幸編)" 広川書店, 148 (1997)

[Publications] 市川一寿: "記憶とは何か記憶の分子生物学(分担執筆)" 丸善, 126 (1997)