種々のレチナールタンパクの光化学反応の相互比較 その機能と構造からの比較

1992 Fiscal Year Annual Research Report

• Project/Area Number: 04302065
• Research Institution: Hokkaido University
• Principal Investigator: 加茂 直樹 北海道大学, 薬学部, 教授 (10001976)
• Co-Investigator(Kenkyū-buntansha): 徳永 史生 大阪大学, 理学部, 教授 (80025452) ; 北川 禎三 岡崎国立共同研究機構分子科学研究所, 教授 (40029955) ; 田隅 三生 東京大学, 理学部, 教授 (60011540) ; 鈴木 英雄 早稲田大学, 理工学部, 教授 (60063541) ; 向畑 恭男 名古屋大学, 理学部, 教授 (10028110)
• Keywords: ロドプシン / バクテリオロドプシン / ハロロドプシン / ホボロドプシン / ゾーリ虫 / クラミドモナス / 高度好塩菌 / 光化学反応

Research Abstract

本研究の目的は、種々のロドプシンおよびロドプシン様タンパク研究者が集まり、討論し、ある場合は共同研究を行ない、ロドプシン(様タンパク)の光化学反応を解明することにある。
1.pharaonis phoborhodopsin(ppR)について:低温スペクトル法、ナノ秒の閃光光分解、ミリ秒閃光光分解法により、ppRの光化学反応を明らかにした。バクテリオロドプシン(bR)の光化学反応と似ているが、M中間体の崩壊の速度が極めて遅い。その速度は外液のpHに依存し、弱酸の陰イオンが加速されることが分かった。
2.halorhodpsin(hR)について:Halobacterium halobium以外からhR様タンパク2種について、それらの一次構造が本研究によって明らかにされた。hRの機能であるClの輸送に必要なアミノ酸残基が明らかになると期待される。2種のうちの1つについては、タンパクも部分精製され、光化学反応が調べられた。
3.bacteriorhodopsin(bR)およびarchearhodopsin(aR)について:アルコール添加により、bRの光化学反応が速くなることを発見した。速くなる処理法の発見は世界で初めてである。ナノ秒の時間分解吸収光度法および閃光光分解吸収異方性測定装置を開発しbRに適用した。aRの明暗順応に必須のアミノ酸残基を同定した。
4.visual pigmentsについて:タコのロドプシンを用いて、ラマン散乱法、赤外吸収法、蛍光ラベル法などにより、構造変化を調べ、新しい中間体を発見した。また、フェムト秒の閃光光分解実験が行なわれた。
5.微生物のロドプシン様タンバクについて:ゾウリ虫では、高等動物と同じようにGタンパクと共役はていること、カエルのロドプシンに対する抗体がクロスすることを示した。クラミドモナスでは、レチナールアナログの実験から、光化学反応の速度が重要であることが分かった。

Research Products

• [Publications] J.Hirayama,: "Photocycle.of phoborhodopsin from haloalkaliphilic bacterium(Natoronobacterium pharaonis)studied by low-temperature spectrophotometry" Biochemistry. 31. 2093-2098 (1992)
• [Publications] Y.Imamoto,: "Chromophore Configuration of pharaonis phoborhodopsin and its isomerization on photon absorption" Biochemistry. 31. 2523-2528 (1992)
• [Publications] M.Miyazaki,: "Flash photolysis study on pharaonis phoborhodpsin from a haloalkaliphilic bacterium(Natronobacterium pharaonis)" Biochim.Biophys.Acta. 1140. 22-29 (1992)
• [Publications] Y.Imamoto,: "Nanosecond laser photolysis of phoborhodopsin from Natronobacterium pharaonis:Appearance of KL and L intermendiates in the photocycle at room temperature" Photochem.Phototobiol. 56. 1129-1134 (1992)
• [Publications] Y.Mukohata,: "Photophosphorylation elements in halobacteria:An A-type ATP synthase and bacterial rhodpsins" J.Bionerg.Biomembr.24. 547-553 (1992)
• [Publications] K.Ihara,: "Halobacterial A-ATP synthase in relation to V-ATPase" J.exp Biol. 172. 475-485 (1992)
• [Publications] Y.Mukohata: "Comparative studies on bacteriorhodopsin-type pumps:Basic physicochemical date for proton pumps" Colloque INSERM. 221. 101-104 (1992)
• [Publications] M.Taiji: "Femtosecond studies of primary photoprocesses in Octopus rhodopsin" Photochem.Photobiol.56. 1003-1011 (1992)
• [Publications] K.Bryl: "Light adaptation of dark-adapted bacteriorhodopsin studied by nanosecond time-resolved absorption spectroscopy" Photochem.Photobiol.56. 1013-1018 (1992)
• [Publications] K.Azuma,: "HPLC analysis of retinoids extracted from the planarian,Dugesia japonica" Zool.Sci.9. 941-946 (1992)
• [Publications] S.Terada,: "Recovery of intact membrane proteins from adherent animal cells grown in a liquid-liquid interface" J.Fermen.Bioenerg.74. 330-332 (1992)
• [Publications] S.A.Salehi,: "Effect of SH reagents on rod photoresponse of isolated frog retina" India J.Pphthalmol.
• [Publications] 寺田 庄一,: "ウシ脳における細胞増殖因子の検索" 北関東医学. 42(2). 77-85 (1992)
• [Publications] H.Kikuchi,: "Dynamical theory of the photoisomerization of rhodopsin chromophore.I.Calculation of the transition probability" J.Phys.Soci.Japan. 61. 1996-1959 (1992)
• [Publications] S.Sato,: "Time-resolved resonance Raman spectra of free-base octaethylporphyrin in the Sl and Tl states" chem.Phys lett. 189. 443-447 (1992)
• [Publications] S.kamikura: "Time-resolved UV resonance Raman study on Bohr effects on hemoglobin" J.Am.Chem.Soc.114. 3256-3260 (1992)
• [Publications] K.Ishimori: "Site-directed mutagenesis in himoglobin:Functional and structural role of intra-and inter-subnuit hydrogen bonds as studied with 37 and 145 mutations" Biochemistry. 31. 3256-3265 (1992)
• [Publications] Y.Sakan: "Detection of the Fe-Co stretching Rama band for recombination transient species of photodissociated carbonmonoxy myoglobin by using pump/probe time resolved technique" chem.Phys.lett.196. 150-154 (1992)
• [Publications] S.Kaminaka: "Resonance Raman study on active site structure of an cooperative hemorythrin" Biochemistry. 31. 6997-7002 (1992)
• [Publications] T.Kitagawa: "Investigation of higher order structures of proteins by ultraviolet resonance Raman spectroscopy" Prog.Biophys.Molec.Biol. 58. 1-18 (1992)
• [Publications] H.Sasabe: "Two-dimensioal molecular packing of proteins" Thin Solid Films. 216. 99-104 (1992)
• [Publications] J.Otomo: "Bacterial rhodopsins of newly isolated halobacteria" I.gen.Microbiol.138. 1027-1037 (1992)
• [Publications] H.Kandori: "Primary Photochemical events in halorhodopsin studied by subpicosecond time-resolved spectroscopy" J.Phys.Chem.96. 6066-6071 (1992)
• [Publications] J.Otomo: "Properties of the primary structure of a new halorhodopsin from halobacterial strain mex" J.Phys.Chem. 96. 6066-6071 (1992)
• [Publications] J.Otomo: "Properties of the primary structure of a new halorhodopsin from halobacterial strain mex" Biochim.Biophys.Acta. 1112. 7-13 (1992)
• [Publications] K.Fukuda,: "Photoreaction of bacteriorhodopsin at high pH:origins of the slow decay component of M" Biochemistry. 31. 11740-11747 (1992)
• [Publications] K.Fukuda,: "Formation and decay kinetics of bacteriorhodopsin's N intermediate:Softening the protein conformation with alcohols affects intra-proton transter and retinal isomerization" Photochem.Photobiol.56. 1057-1062 (1992)
• [Publications] S.Masuda,: "Infrared studies of octopus rhodopsin:Existence of a long-lived intermediate and the states of the carboxylic group of Asp-81 in rhodopsin and its photoroducts" FEBS Letters. 317. 223-227 (1993)
• [Publications] M.kataoka,: "BActeriorhodopsin reconstitued from two individual hilice and an complementary five hilix fragment is photoactive" Photochem.Photobiol.56. 895-890 (1992)
• [Publications] N.Sekiya: "Structural study of retinochrome using fluorinated analogue chromophores" Photochem.Photobiol.56. 909-911 (1992)
• [Publications] T.Iwasa,: "Novel bacteriorhodopsin analogue with conformationally 6-s-cis fixed retinals" Photochem.Photobiol.56. 921-927 (1992)
• [Publications] T.kakitani,: "Excited state dynamic of retinal protein studied by fourie transform of optical absorption spectrum:Development of analytical method" Photoceem.Photobiol. 56. 921-927 (1992)
• [Publications] Y.Beppu,: "Energetics of protonation-deprptpnation of the chromophore in retinal proteins" Photochem.Photobiol.56. 1113-1117 (1992)
• [Publications] Y.Koyama,: "Isomerization of the retinylidene chromophore of bacteriorhodpsin in light adaptation:Intrinsic isomerization of the chromophore and its control by the apo-protein" Photochem.Photobiol.(1993)
• [Publications] M.Nakagawa,: "Identification of the long-lived L'species of bacteriothodopsin tobe the N-intermediates by Raman/absorption quasi-simultaneous measurements" J.Molecul.Struct.242. 221-234 (1991)
• [Publications] T.Okano,: "Cone visual pigments present in gecko rod cells" Proc.Natl.Acad.Scl.USA. 89. 6841-6845 (1992)
• [Publications] M.Ito,: "Conformational analysis of the rhodopsin chromophore using bicyclic retinal analogues" Photochem.Photobiol.56. 915-919 (1992)
• [Publications] J.Sasaki.: "Protein changes associated with reprptonation of the schiff base in photocycle of Asp--Asn Bacteriorhodopsin" J.Biol.Chem.267. 20782-20786 (1992)
• [Publications] Y.Kito,: "Absorbance spectrumand photosensitivity of a new synthetic visual pigment based on 4-hydroxylretinal" Vision Res. 33. 3-10 (1992)
• [Publications] M.Sedou: "Cyclic GMP is not a second messenger in cephalopod retina" Zoologec.Sci.10. (1993)
• [Publications] T.Takahashi,: "Sensitivity increase in the photophobic response of halobacterium halobium reconstituted with retinal anaogs:a novel interpretation for the fluence-response relationship and a kinetic modeling" Photochem.Photobiol.56. 1119-1128 (1992)
• [Publications] N.Sekiya,: "Structural study of retinochrome using fluorinated analog chromophores" Photochem.Photobiol.56. 904-914 (1992)
• [Publications] T.Takahashi,: "Diversion of the sign of phototaxis in a Chlamydomonas reinhardtii mutant incorporated with retinal and its analogs" FEBS lett.314. 275-279 (1992)
• [Publications] 津田 基之: "光センサーロドプシンの機能" 電子写真. 31. 56-61 (1992)
• [Publications] 鬼頭 勇次: "ホタルイカの眼にみる深海生物学" うみうし通信. 8. 22-36 (1992)
• [Publications] 富岡 寛頭: "生物物理(古細菌の走行性の光受容蛋白質)" 生物物理学会, 6 (1992)
• [Publications] H.Tomika,: "Structures and Functions of Retinal Proteins,Ed.J.L.Rigaud(AbSORbance change of a C-50 carotenoid,bacterioruberin related to Cl^- translocation in a Cl^- pump,Halorhodopsin)" John Libbery Eurotext Ltd.Paris, 4 (1992)
• [Publications] Y.Imamoto.: "Structures and Functions of Retinal Proteins,Ed.J.L.Rigaud(Chromophore configuratin and photoreaction cycle of phoborhodopsin from Natronobacterium pharaonis.Structures and Functions of Retinal Proteins)" John Libbery Eurotext Ltd.Paris, 4 (1992)
• [Publications] 藤沢 隆雄: "物理学辞典[改訂版]:タンパク質のリン酸化" 倍風館, 1 (1992)
• [Publications] 小倉 尚志: "新生化学実験講座 第5巻" 東京化学同人, 9 (1992)
• [Publications] 北川 禎三: "生物物理" 生物物理学会(吉岡書店), 5 (1992)
• [Publications] H.Tomioka: "Structures and Functions of Retinal Proteins,Ed.J.L.Rigaud(Absorbance change of a C-50 carotenoid,bacterioruberin related to Cl^- translocation in a Cl^- pump,halorhodopsin)" John Libbery Eurotext Ltd Paris, 4 (1992)
• [Publications] J.Otomo et al.: "Structures and Functions of Retinal Proteins,Ed.J.L.Rigaud(Properties and the primary structures of new bacterial rhodopsins)" John Libbery EUrotext Ltd Paris, 4 (1992)
• [Publications] M.Tsuda: "Structures and Functions of Retinal Petinal Proteins,Ed.J.L.Rigaud" John Libbery Eurotext Ltd Paris, 4 (1992)
• [Publications] M.Tsuda: "11th International congress on Photobiology:Excerpta Medica International Congress Series" Elsevier Science Publishers, 4 (1993)
• [Publications] Y.Imaoto: "Structure and F1nction of Retinal Proteins" Elsevier.The Netherlands, (1993)
• [Publications] Y.Shichida: "Structure and F1nction of Retinal Proteins" Elsevier.The Netherlands, (1993)
• [Publications] M.Seidou,: "Three visual pigments of the firefly souid in Cephalopod neurobiology" Oxford Univ Press, 28 (1992)
• [Publications] Y.Kito: "A blue sensitive new visual pigmen in structure function of retinal proteins" John Libbey Eurotext Ltd, 4 (1992)