1992 年度実績報告書

色受容細胞における分子応答

研究課題

研究課題/領域番号	03454559
研究機関	京都大学
研究代表者	七田芳則京都大学, 理学部, 助手 (60127090)
研究分担者	深田吉孝京都大学, 理学部, 助手 (80165258)
キーワード	視細胞 / ロドプシン / 錐体光受容蛋白質 / G蛋白質 / レーザー閃光分解法 / 低温分光法 / 遺伝子発現 / 情報伝達
研究概要	交付申請書に記載の研究実施計画にそって研究を進め、以下の知見を得た。1.アイオドプシンをサブピコ秒レーザー(波長590nm、パルス幅0.5ピコ秒)で励起した後の吸収変化を測定した。その結果、励起直後にアイオドプシンの励起状態の生成が観測され、その後励起状態は約200フェムト秒の時定数でフォト中間体に変化することがわかった。また、フォト中間体は数ピコ秒の時定数でバソ中間体に変化することがわかった。以上の結果から、アイオドプシンの第一光産物もロドプシンと同様フォト中間体であることがわかった。2.ニワトリ網膜4000枚から青色感受性錐体に存在する光受容蛋白質(ニワトリBlue)を分離・精製することに成功した。また、この試料を用いてその光反応課程を検討したところ、ロドプシンやアイオドプシンと同様、バソ、ルミ、メタ中間体に対応する中間体の生成を確認した。3.メタII中間体とトランスデューシンの相互作用におけるトランスデューシンの翻訳後修飾(ファルネシル化及びメチル化)の生理的役割を明らかにするため、ファルネシル基のみ結合しているトランスデューシンとファルネシル基、メチル基の両方が結合しているものを分離・精製した。それぞれをフォスファチジルコリンのリポソームに組み込んだロドプシンと混合して光照射したところ、メチル化されているトランスデューシンの方がメタII中間体と強く相互作用することがわかった。4.ヒト由来の株細胞である293s細胞中でロドプシンの遺伝子を発現させることに成功した。さらにPCR法を用いて、ロドプシン遺伝子に点変異を導入した変異体を作製し、上記細胞中で発現させることに成功した。

研究成果

(22件)

すべてその他

すべて文献書誌 (22件)

[文献書誌] A.Maeda: "Water structural changes in the bacteriorhodopsin photocycle:Analysis by fourier transform infrared spectroscopy." Biochemistry. 31. 462-467 (1992)
[文献書誌] J.Hirayama: "A photocycle of phoborhodopsin from haloalkaliphilic bacterium(Natronobacterium phoraonis) studied by low-temperature spectrophotometry." Biochemistry. 31. 2093-2098 (1992)
[文献書誌] Y.Imamoto: "Chromophore configuration of pharaonis phoborhodopsin and its isomerization on a photon absorption." Biochemistry. 31. 2523-2528 (1992)
[文献書誌] M.Araki: "Differentiation of both rod and cone types of photoreceptors in the in vivo and in vitro developing pineal glands of the quail." Develop.Brain Res.65. 85-92 (1992)
[文献書誌] T.Okano: "Primary structure of chicken cone visual pigments;Scotopic vision has evolved out of photopic vision." Proc.Natl.Acad.Sci.USA. 89. 5932-5936 (1992)
[文献書誌] D.Kojima: "Cone visual pigments present in gecko rod cells." Proc.Natl.Acad.Sci.USA. 89. 6841-6845 (1992)
[文献書誌] A.Maeda: "Structure of aspartic acid-96 in the L and N intermediates of bacteriorhodopsin;Analysis of fourier transform infrared spectroscopy." Biochemicstry. 31. 4684-4690 (1992)
[文献書誌] M.Ito: "Conformational analysis of the rhodopsin chromophore using bicyclic retinal analogues." Photochem.Photobiol.56. 915-919 (1992)
[文献書誌] T.Kakitani: "Excited state dynamics of retinal proteins as studied by fourier transform of optical absorption spectrum 1.Development of analytical method." Photochem.Photobiol. 56. 977-987 (1992)
[文献書誌] T.Kakitani: "Excited state dynamics of retinal protein as studied by fourier transform of optical spectrum.2.Theoretical analysis of wavepacket propagation of rhodopsin." Photochem.Photobiol.56. 989-993 (1992)
[文献書誌] T.Okano: "Photosensitivities of chicken red rhodopsin" Photochem.Photobiol.56. 995-1001 (1992)
[文献書誌] J.Sasaki: "Stucture of the chromophore in hypsorhodopsin analysed by fourier transform infrared spectroscopy." Photochem.Photobiol.56. 1063-1071 (1992)
[文献書誌] Y.Imamoto: "Nanosecond laser photolysis of pharaonis phoborhodopsin:Appearance of KL and L intermediates in the photocycle at room temperature." Photochem.Photobiol.56. 1129-1134 (1992)
[文献書誌] A.Kawata: "Photoreceptor cell types in the retinal of various vertebrate species:Immunocytochemistry with antibodies against rhodopsin and iodopsin." Photochem.Photobiol.56. 1157-1164 (1992)
[文献書誌] J.Sasaki: "Protein changes associated with reprotonation of the Schiff base in photocycle of Asp^<96>‐‐>Asn bacteriorhodopsin." J.Biol.Chem.267. 20782-20786 (1992)
[文献書誌] T.Mizukami: "Photoisomerization mechanism of the rhodopsin chromophore:Picosecond photolysis of pigment containing 11-cis-locked-8-membered-ring-retinal." Proc.Natl.Acad.Sci.USA. (1993)
[文献書誌] K.Kokame: "Identification of the α-subunits of rod and cone transducin in chicken photoreceptor cells." Exp.Eye Res.(1993)
[文献書誌] A.Wada: "A new rhodopsin analog involving 11Z-8,18-ethanoretinal as a chromophore." Chem.Pharm.Bull.(1993)
[文献書誌] 七田芳則: "色の弁別に関与する視細胞に含まれる光受容蛋白質" 化学と生物. 30. 351-359 (1992)
[文献書誌] 岡野俊行: "錐体視物質の一次構造:ロドプシンは錐体視物質から進化した" 生物物理. 32. 273-276 (1992)
[文献書誌] T.Shichida: "Changes in chromophore-opsin interaction in the photobleaching.processes of visual pigments.“Structures and Functions of Retinal Proteins".Ed.by J.L.Rigaud" John Libbey Eurotext,France, 434 (1992)
[文献書誌] Y.Imamoto: "Chromophore configuration and photoreaction cycle of phobo-rhodopsin from Natronobacterium pharaonis.“Structures and Functions of Retinal Proteins".Ed.by J.L.Rigaud." John Libbey Eurotext,France, 434 (1992)

1992 年度 実績報告書

色受容細胞における分子応答

研究代表者

七田 芳則 京都大学, 理学部, 助手 (60127090)

研究成果

[文献書誌] A.Maeda: "Water structural changes in the bacteriorhodopsin photocycle:Analysis by fourier transform infrared spectroscopy." Biochemistry. 31. 462-467 (1992)

[文献書誌] J.Hirayama: "A photocycle of phoborhodopsin from haloalkaliphilic bacterium(Natronobacterium phoraonis) studied by low-temperature spectrophotometry." Biochemistry. 31. 2093-2098 (1992)

[文献書誌] Y.Imamoto: "Chromophore configuration of pharaonis phoborhodopsin and its isomerization on a photon absorption." Biochemistry. 31. 2523-2528 (1992)

[文献書誌] M.Araki: "Differentiation of both rod and cone types of photoreceptors in the in vivo and in vitro developing pineal glands of the quail." Develop.Brain Res.65. 85-92 (1992)

[文献書誌] T.Okano: "Primary structure of chicken cone visual pigments;Scotopic vision has evolved out of photopic vision." Proc.Natl.Acad.Sci.USA. 89. 5932-5936 (1992)

[文献書誌] D.Kojima: "Cone visual pigments present in gecko rod cells." Proc.Natl.Acad.Sci.USA. 89. 6841-6845 (1992)

[文献書誌] A.Maeda: "Structure of aspartic acid-96 in the L and N intermediates of bacteriorhodopsin;Analysis of fourier transform infrared spectroscopy." Biochemicstry. 31. 4684-4690 (1992)

[文献書誌] M.Ito: "Conformational analysis of the rhodopsin chromophore using bicyclic retinal analogues." Photochem.Photobiol.56. 915-919 (1992)

[文献書誌] T.Kakitani: "Excited state dynamics of retinal proteins as studied by fourier transform of optical absorption spectrum 1.Development of analytical method." Photochem.Photobiol. 56. 977-987 (1992)

[文献書誌] T.Kakitani: "Excited state dynamics of retinal protein as studied by fourier transform of optical spectrum.2.Theoretical analysis of wavepacket propagation of rhodopsin." Photochem.Photobiol.56. 989-993 (1992)

[文献書誌] T.Okano: "Photosensitivities of chicken red rhodopsin" Photochem.Photobiol.56. 995-1001 (1992)

[文献書誌] J.Sasaki: "Stucture of the chromophore in hypsorhodopsin analysed by fourier transform infrared spectroscopy." Photochem.Photobiol.56. 1063-1071 (1992)

[文献書誌] Y.Imamoto: "Nanosecond laser photolysis of pharaonis phoborhodopsin:Appearance of KL and L intermediates in the photocycle at room temperature." Photochem.Photobiol.56. 1129-1134 (1992)

[文献書誌] A.Kawata: "Photoreceptor cell types in the retinal of various vertebrate species:Immunocytochemistry with antibodies against rhodopsin and iodopsin." Photochem.Photobiol.56. 1157-1164 (1992)

[文献書誌] J.Sasaki: "Protein changes associated with reprotonation of the Schiff base in photocycle of Asp^<96>‐‐>Asn bacteriorhodopsin." J.Biol.Chem.267. 20782-20786 (1992)

[文献書誌] T.Mizukami: "Photoisomerization mechanism of the rhodopsin chromophore:Picosecond photolysis of pigment containing 11-cis-locked-8-membered-ring-retinal." Proc.Natl.Acad.Sci.USA. (1993)

[文献書誌] K.Kokame: "Identification of the α-subunits of rod and cone transducin in chicken photoreceptor cells." Exp.Eye Res.(1993)

[文献書誌] A.Wada: "A new rhodopsin analog involving 11Z-8,18-ethanoretinal as a chromophore." Chem.Pharm.Bull.(1993)

[文献書誌] 七田 芳則: "色の弁別に関与する視細胞に含まれる光受容蛋白質" 化学と生物. 30. 351-359 (1992)

[文献書誌] 岡野 俊行: "錐体視物質の一次構造:ロドプシンは錐体視物質から進化した" 生物物理. 32. 273-276 (1992)

[文献書誌] T.Shichida: "Changes in chromophore-opsin interaction in the photobleaching.processes of visual pigments.“Structures and Functions of Retinal Proteins".Ed.by J.L.Rigaud" John Libbey Eurotext,France, 434 (1992)

[文献書誌] Y.Imamoto: "Chromophore configuration and photoreaction cycle of phobo-rhodopsin from Natronobacterium pharaonis.“Structures and Functions of Retinal Proteins".Ed.by J.L.Rigaud." John Libbey Eurotext,France, 434 (1992)

1992 年度実績報告書

七田芳則京都大学, 理学部, 助手 (60127090)

[文献書誌] 七田芳則: "色の弁別に関与する視細胞に含まれる光受容蛋白質" 化学と生物. 30. 351-359 (1992)

[文献書誌] 岡野俊行: "錐体視物質の一次構造:ロドプシンは錐体視物質から進化した" 生物物理. 32. 273-276 (1992)