光合成系におけるカロテノイドの機能とその基礎としての反応動力学の解析

1990 Fiscal Year Annual Research Report

• Project/Area Number: 02305014
• Research Institution: Okazaki National Research Institutes
• Principal Investigator: 三室 守 岡崎国立共同研究機構, 基礎生物学研究所, 助手 (40142004)
• Co-Investigator(Kenkyū-buntansha): 垣谷 俊昭 名古屋大学, 理学部, 助教授 (90027350) ; 山崎 巌 北海道大学, 工学部, 教授 (80002111) ; 加藤 哲也 京都大学, 理学部, 講師 (30025308) ; 小山 泰 関西学院大学, 理学部, 教授 (90079666)
• Keywords: カロテノイド / 光合成 / エネルギ-移動 / 光合成色素蛋白 / 時間分解分光法 / 分子動力学

Research Abstract

1.光合成色素カロテノイドの機能の中で、特に光捕獲アンテナ色素としての機能、すなわち一重項ー一重項エネルギ-転移機構を、生化学的、分光学的、分子動力学的に解析し、多くの成果を得た。
2.褐藻、珪藻、渦鞭毛藻などから、エネルギ-転移機能を保持した色素蛋白複合体の分離精製方法を確立し、さらに複合体が黄色植物に広く分布することを明らかにした。次に大量調製法を確立した。これにより、共通の試料について多方面からの解析が可能となり、大きな進展につながった。
3.エネルギ-転移過程の分光学的解析から、エネルギ-転移に関与する準位が、可視部の吸収に対応する第二励起準位ではなく、一光子過程としては禁制遷移の第一励起準位を介して起こること、その寿命が10ー40ピコ秒(種によって異なる)であることを明らかにした。
4.この過程の理論的解析の第一歩として、分子軌道法を採用し、<in vitro>___ーでの分子構造の最適化のためのパラメトリゼ-ションを行った。ついで、炭化水素のみから構成されるカロテノイドからクロロフィルへのエネルギ-転移過程を、従来考えられていた電子交換相互作用ではなく、ク-ロン相互作用(四極子ー双極子相互作用)での可能性を検討し、特定の配向では十分な速度が期待されることを明らかにした。
5.反応機構に密接に関係するカロテノイドの物性を知るため、色素分析法の確立、構造解析法の改良を行なった。これを応用して、光合成細菌から新しい分子種の単離し、その構造決定に成功した。藻類のカロテノイド蛋白質の構造を、部分的な一次構造の決定や、hydropathy plotにより解析した。
6.三重項ー三重項遷移過程については、今年度は実質的な解析ができず、来年度の検討課題となった。

Research Products

• [Publications] Mimuro,M.et al.,: "Spatial arrangement of pigments and their interaction in the fucoxanthinchl a__ー/c__ー protein assembly (FCPA) isolated from the brown alga <Dictyota>___ー <dichotoma>___ー:Analysis by means of polarized spectroscopy." Biochim.Biophys.Acta,. 1015. 450-456 (1990)
• [Publications] Mimuro,M.et al.,: "Characteristic fluorescence components in the pigment system of the marine dinoflagellate <Protogonyaulax tamarensis>___ー and energy flow among them:Studies by means of steadyーstate and timeーresolved fluorescence spectroscopy." Biochim.Biophys.Acta,. 1016. 281-287 (1990)
• [Publications] Mimuro,M.: "Excitation energy flow in the photosynthetic pigment systems:structure and energy transfer mechanism." Bot.Mag.Tokyo,. 103. 233-253 (1990)
• [Publications] Miki,K.et al.,: "Crystallization and preliminary Xーray diffraction studies of Cーphycocyanin from a red alga <Porphyra tenera>___ー." J.Biochem.(Tokyo),. 108. 646-649 (1990)
• [Publications] Nozawa,T.et al.,: "Magnetic circular dichroism and linear dichrosim for elucidation of electronic transitions and their orientations in a reaction center isolated from a thermophilic purple sulfur photosynthetic bacterium <Chromatium>___ー <tepidum>___ー." Chem.Lett.,. 2125-2128 (1990)
• [Publications] 三室 守: "アンテナ複合体における分子配置" 細胞. 22. 296-300 (1990)
• [Publications] Mimuro,M.et al.,: "Carotenoids in photosynthesisーAbsorption,transfer and dissipation of light energy." Pure & Appl.Chem.,. 63. 123-130 (1991)
• [Publications] Uehara,K.et al.,: "Spectroscopic studies of chlorophyll a__ー aggregates formed by aqueous Dimethyl sulfoxide." Photochem.Photobiol.,.
• [Publications] Frackowiak,D.et al.,: "Analysis of timeーresolved emission spectra of oriented phycobilisomes." Photochem.Photobiol.,.
• [Publications] Mimuro,M.et al.,: "Fluorescence properties of the allenic carotenoid fucoxanthin:Analysis of the effect of keto carbonyl group by using a model compound,allーtransーβーapoー8'ーcarotenal." J.Luminescence.
• [Publications] 三室 守: "フェムト秒の生物学 光合成の物理化学的側面" パリティ-.
• [Publications] Yoshimori,A.et al.,: "Theoretical study of the free energy curves as a function of Reaction coordinate of electron transfer by the cumulant expansion method." J.Chem.Phys.93. 5140-5146 (1990)
• [Publications] Beppu,Y.et al.,: "Comprehensive study of molecular conformation and adiabatic potential surface of allーtrans retinal by the CNDO/2 method." Chem.Phys.148. 333-346 (1990)
• [Publications] Saito,M.et al.,: "Monte Carlo simulation study on solvation energy of polar solutions." Chem.Phys.Lett.172. 169-174 (1990)
• [Publications] Hatano,Y.et al.,: "Theoretical Study of Dielectric Saturation in Molecular solutions by the Monte Carlo Simulation." J.Phys.Soc.Japan. 59. 1104-1116 (1990)
• [Publications] Suzuki,H.et al.,: "An extended INDOーCI study on the interaction of formamide with exteral electric field." J.Phys.Soc.Japan. 59. 3393-3400 (1990)
• [Publications] Sugimoto,T.et al.,: "An extended INDOーCI study on protonated retinal Schiffーbase." J.Phys.Soc.Japan. 59. 3780-3790 (1990)
• [Publications] Kim,I.H.et al.,: "Structure and function of the photoreceptor stentorins in <Stentor>___ー coeruleus.I.Partial characterization of the photoreceptor organella and stentorins." Biochim.Biophys.Acta. 1040. 43-57 (1990)
• [Publications] Song,P.S.et al.,: "Structure and Function of the photoreceptor stentorins in <Stentor>___ー coeruleus.II.Primary photoprocess and picosecond timeーresolved fluorescence." Biochim.Biophys.Acta. 1040. 58-65 (1990)
• [Publications] Kuki,M.et al.,: "The 2^1Ag^ー state of a carotenoid bound to the chromatophore membrane of <Rhodobacter sphaeroides>___ー 2.4.1 as revealed by transient resonance Raman spectroscopy." Chem.Phys.Lett.165. 417-422 (1990)
• [Publications] Koyama,Y.et al.,: "Configurations of Carotenoids in the reaction center and the lightーharvesting complex of <Rhodospirillum rubrum>___ー.Natural selection of carotenoid configurations by pigment protein complexes." Photochem.Photobiol.51. 119-128 (1990)
• [Publications] Mukai,Y.et al.,: "Dependence of the triplet potential of retinal homologs on the chain length:Resonance Raman spectroscopy and analysis of tripletーsensitized isomerization." J.Phys.Chem.94. 4042-4051 (1990)
• [Publications] Hashimoto,H.et al.,: "The 2^1Ag^ー state of a carotenoid bound to spinach chloroplast as revealed by picosecond transient Raman spectroscopy." Biochim.Biophys.Acta. 1017. 181-186 (1990)
• [Publications] Nishizawa,E.et al.,: "The T_1 Raman spectra of bacteriochlorophyll a__ー:Changes in the bond orders of the conjugated macrocycle upon triplet excitation." Chem.Phys.Lett.172. 317-322 (1990)
• [Publications] Katayama,N.et al.,: "Highーperformance liquid chromatography of <cisーtrans>___ー isomers of neurosporene:Discrimination of <cis>___ー and <trans>___ー configurations at the end of an open conjugated chain." J.Chromatogr.519. 221-227 (1990)
• [Publications] Naruse,M.et al.,: "Triplet excitation of precursors of spirilloxanthin bound to the chromatophores of <Rhodospirillum rubrum>___ー as detected by transient Raman spectroscopy." J.Mol.Struct.242. 15-26 (1991)
• [Publications] Nishizawa,E.et al.,: "A Coordinationーmarker Raman line of bacteriochlorophyll a__ー in the T_1 state." Chem.Phys.Lett.176. 390-394 (1991)
• [Publications] Hayashi,H.et al.,: "Picosecond timeーresolved resonance Raman scattering and absorbance changes of carotenoids in light harvesting systems of the photosynthetic bacterium <Chromatium vinosum>___ー." J.Am.Chem.Soc.112. 4664-4670 (1990)
• [Publications] Noguchi,T.et al.,: "Factors controlling the efficiency of energy transfer from carotenoids to bacteriochlorophyll in purple photosynthetic bacteria." Biochim.Biophys.Acta. 1017. 280-290 (1990)
• [Publications] Sakai,J.et al.,: "Infrared studies of the phototransformation of phytochrome." Chem.Lett.1925-1926 (1990)
• [Publications] Noguchi,T.et al.,: "Frequencies of the FrankーCondon active a_g C=C streching mode in the 2^1A_g^ー states of carotenoids." Chem.Phys.Lett.(1990)
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• [Publications] Takaichi,S.et al.,: "Carotenoids from the aerobic photosynthetic bacterium,<Erythrobacter>___ー <longus>___ー:βーCarotene and its hydroxyl derivatives." Arch.Microbiol.153. 118-122 (1990)
• [Publications] Takaichi,S.et al.,: "Caroenoid pigments from <Rhodococcus rhodochrous>___ー RNMS1:Two monocyclic carotenoids,a carotenoid monoglycoside and carotenoid glycoside monoesters." Agric.Biol.Chem.54. 1931-1937 (1990)
• [Publications] Takaichi,S.: "Heterogeneous position of the double bonds of unsaturated fatty acids in carotenoid glucoside esters from <Rhodoccus rhodochrous>___ー RNMS1." Agric.Biol.Chem.54. 2139-2140 (1990)
• [Publications] Takaichi,S.et al.,: "Light induced changes of carotenoid pigments in anaerobic cells of the aerobic photosynthetic bacterium,<Roseobacter denitrificans>___ー (<Erythrobacter>___ー species OCh 114):Reduction of spheroidenone to 3,4ーdihydrospheroidenone." Arch.Microbiol.
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• [Publications] Katoh,T.et al.,: "Supramolecular assembly of fucoxanthinーchlorophyllーprotein complexes isolated from a brown alga,<Petalonia fascia>___ー.Electron microscopic studies." Plant Cell Physiol.31. 439-447 (1990)
• [Publications] 加藤 哲也: "褐藻の光捕捉顆粒" 海洋. 22. 717-721 (1990)
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• [Publications] Shimada,K.et al.,: "Excitation energy flow in a photosyntheticbascterium laking B850.Fast energy flow from B806 to B870 in <Erythrobacter>___ー sp.strain OCh 114." Biochim.Biophys.Acta. 1016. 266-271 (1990)
• [Publications] Shimada,K.et al.,: "Excitation and emission spectroscopy of membranes and pigmentーprotain complexes of an aerobic photosynthetic bacterium <Erythrobacter>___ー sp.OCh 114." Plant Cell Physiol.31. 395-398 (1990)
• [Publications] Ogata,T.et al.,: "Peridininーchlorophyll a__ー protein contents of dinoflagelattes <Alexandrium tamarense>___ー and <catenella>___ー." J.Appl.Phycol.
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• [Publications] Mimuro,M.et al.,: "Molecular Biology of MembraneーBound Complexes in Phototrophic Bacteria." Plenum Press, (1990)
• [Publications] 三室 守: "分光技法ハンドブック III 応用編(南 茂夫、合志 陽一編集)" 朝倉図書, (1990)
• [Publications] 垣谷 俊昭: "生物物理の最前線 日本生物物理学会編" 講談社, 302 (1990)
• [Publications] Koyama,Y.: "Carotenoids:Chemistry and Biology" Plenum Press, (1990)
• [Publications] Takaichi,S.et al.,: "Methods in Enzymology" Academic Press,
• [Publications] Takaichi,S.: "Methods in Enzymology" Academic Press,
• [Publications] Katoh,T.et al.,: "Methods in Enzymology:Carotenoids" Academic Press,