1992 Fiscal Year Annual Research Report

色受容細胞における分子応答

Research Project

Project/Area Number	03454559
Research Institution	Kyoto University
Principal Investigator	七田芳則京都大学, 理学部, 助手 (60127090)
Co-Investigator(Kenkyū-buntansha)	深田吉孝京都大学, 理学部, 助手 (80165258)
Keywords	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
Research Abstract	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。

Research Products
(22 results)

All Other

All Publications (22 results)

[Publications] A.Maeda: "Water structural changes in the bacteriorhodopsin photocycle:Analysis by fourier transform infrared spectroscopy." Biochemistry. 31. 462-467 (1992)
[Publications] J.Hirayama: "A photocycle of phoborhodopsin from haloalkaliphilic bacterium(Natronobacterium phoraonis) studied by low-temperature spectrophotometry." Biochemistry. 31. 2093-2098 (1992)
[Publications] Y.Imamoto: "Chromophore configuration of pharaonis phoborhodopsin and its isomerization on a photon absorption." Biochemistry. 31. 2523-2528 (1992)
[Publications] 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)
[Publications] 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)
[Publications] D.Kojima: "Cone visual pigments present in gecko rod cells." Proc.Natl.Acad.Sci.USA. 89. 6841-6845 (1992)
[Publications] 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)
[Publications] M.Ito: "Conformational analysis of the rhodopsin chromophore using bicyclic retinal analogues." Photochem.Photobiol.56. 915-919 (1992)
[Publications] 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)
[Publications] 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)
[Publications] T.Okano: "Photosensitivities of chicken red rhodopsin" Photochem.Photobiol.56. 995-1001 (1992)
[Publications] J.Sasaki: "Stucture of the chromophore in hypsorhodopsin analysed by fourier transform infrared spectroscopy." Photochem.Photobiol.56. 1063-1071 (1992)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] K.Kokame: "Identification of the α-subunits of rod and cone transducin in chicken photoreceptor cells." Exp.Eye Res.(1993)
[Publications] A.Wada: "A new rhodopsin analog involving 11Z-8,18-ethanoretinal as a chromophore." Chem.Pharm.Bull.(1993)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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 Fiscal Year Annual Research Report

色受容細胞における分子応答

Principal Investigator

七田 芳則 京都大学, 理学部, 助手 (60127090)

Research Products

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

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

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

[Publications] 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)

[Publications] 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)

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

[Publications] 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)

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

[Publications] 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)

[Publications] 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)

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

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

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

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

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

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

七田芳則京都大学, 理学部, 助手 (60127090)

[Publications] 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)

[Publications] 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)