オルガネラ内タンパク質の分子科学

• Project/Area Number: 20050029
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: University of Hyogo
• Principal Investigator: 小倉 尚志 University of Hyogo, 大学院・生命理学研究科, 教授 (70183770)
• Project Period (FY): 2008 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥3,800,000 (Direct Cost: ¥3,800,000); Fiscal Year 2009: ¥1,900,000 (Direct Cost: ¥1,900,000); Fiscal Year 2008: ¥1,900,000 (Direct Cost: ¥1,900,000)
• Keywords: プロトンポンプ / チトクロムc酸化酵素 / 酸素活性化 / 共鳴ラマン / 生体エネルギー変換 / 再構成リン脂質小胞 / インドールアミンジオキシゲナーゼ / フェリルオキソ型反応中間体 / ミトコシドリア / 酸化的リン酸化 / 呼吸調節 / プロポンプ / 電子伝達系 / 再構成小胞

Research Abstract

1. 共鳴ラマンスペクトルの測定に適しだチトクロムc酸化酵素再構成ベシクル(COV)の作成法を検討した。CcOとリン脂質の量比、COV調製後の濃縮の方法などが検討課題であり、最終標品の量、呼吸調節率および共鳴ラマンスペクトルの質を指標に最適な手順を決定した。再構成時に脂質吸着ビーズを添加すると、レーザー光に対する耐性が高まることがはっきりした。また、この試料に対し、プロトン濃度勾配存在下での共鳴ラマンスペクトルを測定した。2.インドールアミン-2,3-ジオキシゲナーゼ(IDO)はヘム酵素であり、トリプトファン(Trp)に酸素分子由来の2原子の酸素を添加し、N-フォルミルキヌレニンを生成する反応を触媒する。哺乳類においてこの反応はTrp異化反応の主要経路にある。近年、Trpの代謝産物が免疫系や神経系などの異常に関係することが明らかになり、注目されている。IDOによる酸素添加反応は、従来、Fe-O-O-Trp型の反応中間体を経てO-O結合が切れることなく取り込まれると考えられてきた。本研究では、酸素化型および593nmに吸収帯を持つ新規スペクトル種の共鳴ラマンスペクトルを測定し、これら2つの反応中間体の配位構造を調べた。酸素化型の共鳴ラマンスペクトルより、酸素化型反応中間体がV_<Fe-O2>(鉄-酸素伸縮振動)モードを569cm^<-1>に示し、エンドオン型(Fe-O-O)であることがわかった。一方、酸素化型にTrpを加えて得られる593nmスペクトル種は<16>^O_2に対し798cm^<-1>にラマン線を与えたが、これは<18>^O_2では762cm^<-1>に、また<16>^O<18>^Oに対しては798cm^<-1>と762cm^<-1>に50%ずつの強度のラマン線を与えた。以上の結果から、このものがFe=O型ヘムを持つことが明らかになった。この結果は、従来提唱されてきた反応機構の再考をせまるものである。

Research Products

• [Journal Article] Resonance Raman study on the oxygenated and the ferryl-oxo species of indoleamine 2, 3-dioxygenase during catalytic turnover (2010)
  • Author(s): Sachiko Yanagisawa, Masaki Horitani, Hiroshi Sugimoto, Yoshitsugu Shiro, Norihiro Okada, Takashi Ogura
  • Journal Title: Faraday Discussions (印刷中)
  • Peer Reviewed
• [Journal Article] Cell-free synthesis of cytochrome c oxidase, a multicomponent membrane protein (2010)
  • Author(s): Yukie Katayama, Kunitoshi Shimokata, Makoto Suematsu, Takashi Ogura, Tomitake Tsukihara, Shinya Yoshikawa, Hideo Shimada
  • Journal Title: J.Bioenergetics and Biomembranes
  • Peer Reviewed
• [Journal Article] A Resonance Raman Band Assignable to the O-O Stretching Mode inthe Resting Oxidized State of Bovine Heart Cytochrome C Oxidase (2010)
  • Author(s): Miyuki Sakaguchi, Kyoko Shinzawa-Itoh, Shinya Yoshikawa, Takashi Ogura
  • Journal Title: J.Bioenergetics and Biomembranes
  • Peer Reviewed
• [Journal Article] Nickel(II) complexes of TPA ligands with 6-phenyl substituents PhnTPA. Structure and H_2O_2-reactivity (2010)
  • Author(s): Tano, T., Doi, Y., Inosako, M., Kunishita, A., Kubo, M., Ishimaru, H., Ogura, T., Sugimoto, H., Itoh, S.
  • Journal Title: Bull.Chem.Soc.Jpn. (印刷中)
  • Peer Reviewed
• [Journal Article] A square-planar Ni(II) complex with an N_2S_2 donor set similar to the active centre of nickel-containing superoxide dismutase and its reaction with superoxide (2010)
  • Author(s): Nakane, D., Kuwasako, S., Tsuge, M., Kubo, M., Funahashi, Y., Ozawa, T., Ogura, T., Masuda, H.
  • Journal Title: Chem.Commun. Pages: 2142-2144
  • Peer Reviewed
• [Journal Article] Resonance Raman Study of a High-valent Fe=O Porphyrin Complex as a Model for Peroxide Compound II (2010)
  • Author(s): Hirohito Ishimaru, Hiroshi Fujii, Takashi Ogura
  • Journal Title: Chem.Lett. 39 Pages: 332-333
  • Peer Reviewed
• [Journal Article] Identification of the Fe-O_2 and the Fe=O Heme Species for Indoleamine 2, 3-Dioxygenase during Catalytic Turnover (2010)
  • Author(s): Yanagisawa, S., Yotsuya, K., Hashiwaki, Y., Horitani, M., Sugimoto, H., Shiro, Y., Appelman, E.H., Ogura
  • Journal Title: Chem.Lett. 39 Pages: 36-37
  • NAID: 10027176398
  • Peer Reviewed
• [Journal Article] Spectroscopic Characterization of a Hydroperoxo-Heme Intermediate : Conversion of a Side-On Peroxo to an End-On Hydroperoxo Complex (2009)
  • Author(s): Liu, J.G., Ohta, T., Yamaguchi, S., Ogura, T., .Sakamoto, S., Maeda, Y., Naruta, Y.
  • Journal Title: Angew.Chem.Int.Ed. 48 Pages: 9262-9267
  • Peer Reviewed
• [Journal Article] Geometric and electronic structure and reactivity of a mononuclear 'side-on' nickel(III)- (2009)
  • Author(s): Cho, J., Sarangi, R., Annaraj, J., Kim, S.Y., Kubo, M., Ogura, T., Solomon, E.I., Nam. W.
  • Journal Title: Nature Chemistry 1 Pages: 568-572
  • Peer Reviewed
• [Journal Article] Oxidation Reactivity of a Bis(μ-oxo) Dinickel(III) Complexes : Arene Hydroxylation of the Supporting Ligand (2009)
  • Author(s): Honda, K., Cho, J., Matsumoto, T.,Roh, J., Furutachi, H,, Tosha, T., Kubo, M., Fujinami, S., Ogura, T., Kitagawa, T., Suzuki, M.
  • Journal Title: Angewandte Chemie, Int.Ed. 121 Pages: 3354-3357
  • Peer Reviewed
• [Journal Article] Mononuclear Copper (II)-Superoxo Complexes that Mimic the Stucture and Reactivity of the Active Centers of PHM and DβM (2009)
  • Author(s): Kunishita, A., Kubo, M., Suggimoto, H., Ogura, T., Sato, K., Takui, T., Itoh, S.
  • Journal Title: J.Am.Chem.Soc. 131 Pages: 2788-2789
  • Peer Reviewed
• [Journal Article] Cooperative Binding of L-Trp to Human Tryptophan 2, 3-Dioxygenase : Resonance Raman Spectroscopic Analysis (2009)
  • Author(s): Eiko Fukumura, et.al.
  • Journal Title: Journal of Biochemistry 145 Pages: 505-515
  • Peer Reviewed
• [Journal Article] A peroxide bridge between Fe and Cu ions in the O_2 reduction site of fully oxidized cytochrome c oxidase could suppress the proton pump (2009)
  • Author(s): Hiroshi Aoyama, et.al.
  • Journal Title: Proc. Natl. Acad. Sci. U. S. A. 106 Pages: 2165-2169
  • Peer Reviewed
• [Journal Article] Red-Excitation Resonance Raman Analysis of the VFe=o Mode of Ferryl-Oxo Hemoproteins (2008)
  • Author(s): Kenichiro Ikemura, et.al.
  • Journal Title: J. Am. Chem. Soc. 130 Pages: 14384-14385
  • Peer Reviewed
• [Presentation] 共鳴ラマン分光法によるウシ心筋ミトコンドリア呼吸鎖NADH : ユビキノン酸化還元酵素のFMNと鉄-硫黄クラスターの構造解析 (2010)
  • Author(s): 引田理英、森山昌和、小倉尚志、伊藤-新澤恭子、吉川信也
  • Organizer: 第36回生体分子科学討論会
  • Place of Presentation: 札幌
• [Presentation] 共鳴ラマン分光法によるインドールアミン2,3-ジオキシナーゼの反応中間体の構造解析 (2010)
  • Author(s): 柳澤幸子、四ッ谷景子、橋脇裕美、堀谷正樹、杉本宏、城宜嗣、小倉尚志
  • Organizer: 第36回生体分子科学討論会
  • Place of Presentation: 札幌
• [Presentation] 代謝回転中インドールアミン2,3ジオキシゲナーゼの酸素化型およびフェリルオキソ種の検出 (2010)
  • Author(s): 柳澤幸子・堀谷正樹・杉本宏・城宜嗣・Appelman, Evan H.・小倉尚志
  • Organizer: 日本化学会第90春季年会
  • Place of Presentation: 東大阪
• [Presentation] 高酸化型鉄ポルフィリンの共鳴ラマンスペクトル:Fe=O伸縮振動数の軸配位子依存性 (2010)
  • Author(s): 石丸裕士・藤井浩・小倉尚志
  • Organizer: 日本化学会第90春季年会
  • Place of Presentation: 東大阪
• [Presentation] Structure and Dynamics of Cytochrome c Oxidase as Studiedwith Resonance Raman Spectroscopy (2009)
  • Author(s): Takashi Ogura
  • Organizer: The 14tb International Conference on Biological Inorganic Chemistry (招待講演)
  • Place of Presentation: Nagoya
• [Presentation] Time-Resolved Resonance Raman Study on Structural Relaxation Process of Cytochrome c Oxidase Following Photolysis of Carbonmonoxide (2009)
  • Author(s): Izumi Ishigami, Satoru Nakashima, Kyoko Shinzawa-Itoh, Shinya Yoahikawa, Takashi Ogura
  • Organizer: The 14^<th> International Conference on Bioinorganic Chemistry
  • Place of Presentation: Nagoya
• [Presentation] A New Resonance Raman Marker Band of Cytochrome c Oxidase (2009)
  • Author(s): Miyuki Sakaguchi, Kyoko Shinzawa-Itoh, Shinya Yoshikawa, Hiroshi Fuiii, Takashi Ogura
  • Organizer: The 14^<th> International Conference on Bioinorganic Chemistry
  • Place of Presentation: Nagoya
• [Presentation] X-ray Structure and Function of Peroxide Bridge between Feand Cu in the O_2 Reduction Site of the Fully Oxidized Cytochrome c Oxidase (2009)
  • Author(s): Hiroshi Aoyama, Kazumasa Muramoto, Kyoko Shinzawa-Itoh, Kunio Hirata, Eiki Yamashita, Tomitake Taukihara, Takashi Ogura, Shinya Yoshikawa
  • Organizer: The 14^<th> International Conference on Bioinorganic Chemistry
  • Place of Presentation: Nagoya
• [Presentation] Reactivity of Ru(IV)-Oxo Complexes Derived from Proton-Coupled Electron Transfer of Ru(II)-Aqua Complexes (2009)
  • Author(s): Takahiro Kojima, Yuichiro Hirai, Yasuhisa Mizutani, Kenichiro Ikemura, Takashi Ogura, Yoahihito Shiota, Kazunari Yoshizawa, Shunichi Fukuzumi
  • Organizer: The 14^<th> International Conference on Bioinorganic Chemistry
  • Place of Presentation: Nagoya
• [Presentation] Accurate Design of Copper-containing Oxidase Active Centers with a S-containing Ligand (2009)
  • Author(s): Takashi Tozuka, Yuji Kajita, Minoru Kubo, Takashi Ogura, Yasuhiro Funahashi, Tomohiro Ozawa, Hideki Masuda
  • Organizer: The 14^<th> International Conference on Bioinorganic Chemistry
  • Place of Presentation: Nagoya
• [Presentation] Structure and Reactivity of Mononuclear Copper(II)-Superoxo Complexes Supported by N_3-Tridentate Ligands (2009)
  • Author(s): Atsushi Kunishita, Hideki Sugimoto, Minoru Kubo, Takashi Ogura, Takeji Takui, Shinobu Itoh
  • Organizer: The 14^<th> International Conference on Bioinorganic Chemistry
  • Place of Presentation: Nagoya
• [Presentation] Preparation of Low-spin End-on Ferric-Peroxo and Ferric-Hydroperoxo Species by a Heme Model Complex Bearing an Imidazole Tail (2009)
  • Author(s): Jin-Gang Liu, Takehiro Ohta, Satoru Yamaguchi, Takashi Ogura, Yoahinori Naruta
  • Organizer: The 14^<th> International Conference on Bioinorganic Chemistry
  • Place of Presentation: Nagoya
• [Presentation] Detection of the Iron-Oxygen Stretching Raman Mode for Human Indoleamine 2, 3-Dioxygenase in the Oxygenated Form (2009)
  • Author(s): Sachiko Yanagisawa, Keiko Yotauya, Masaki Horitani, Hiroahi Sugimoto, Yoshitsugu Shiro, Yumi Hashiwaki, Takashi Ogura
  • Organizer: The 14^<th> International Conference on Bioinorganic Chemistry
  • Place of Presentation: Nagoya
• [Presentation] C-H Bond Activation by Peroxodiiron(III) Complexes (2009)
  • Author(s): Susumu Kyuda, Yoshimi Kawade, Hideki Furutachi, Shuhei Fujinami, Masatatsu Suzuki, Takehiko Tosha, Teizo Kitaaggaawa, Minoru Kubo, Takaslri Ocura
  • Organizer: The 14^<th> International Conference on Bioinorganic Chemistry
  • Place of Presentation: Nagoya
• [Presentation] Contribution of the Supporting Ligands to NiO_2 Intermediates (2009)
  • Author(s): Jaeheung Cho, Ritimukta Sarangi, Jameaponck Annaraj, Sung Yeon Kim, Minoru Kubo, Takashi Ogura, Edward I. Solomon, Wonwoo Nam
  • Organizer: The 14^<th> International Conference on Bioinorganic Chemistry
  • Place of Presentation: Nagoya
• [Presentation] Cooperative L-Trp Binding to Human Tryptophan 2, 3-Dioxygenase Revealed by Resonance Raman Spectroscopy (2009)
  • Author(s): Eiko Fukumura, Hiroshi Sugimoto, Yuko Misumi, Takashi Ogura, Yoehitsugu Shiro
  • Organizer: The 14^<th> International Conference on Bioinorganic Chemistry
  • Place of Presentation: Nagoya
• [Presentation] Detection of the Iron-Oxygen Stretching Raman Mode for Human Indoleamine 2, 3-Dioxygenase in the Oxygenated Form (2009)
  • Author(s): Sachiko Yanagisawa
  • Organizer: International Workshop on Metalloprotein Functiona
  • Place of Presentation: Kamigori, Hyogo
• [Presentation] Ultraviolet resonance Raman epectroscopy for functioning proteins (2009)
  • Author(s): Takashi Ogura
  • Organizer: 第47回生物物理学会年会(招待講演)
  • Place of Presentation: 徳島
• [Presentation] Site-directed isotope labeling of cytochrome c oxidase (2009)
  • Author(s): Hideo Shimada, Yukie Katayama, Takashi Ogura, Tomitake Tsukihara, Shinya Yoahikawa
  • Organizer: 第47回生物物理学会年会(招待講演)
  • Place of Presentation: 徳島
• [Presentation] Resonance Raman Marker_ Bands of Hydroxyfarnesylethyl Substituent of Cytochrome c Oxidase (2009)
  • Author(s): Miyuki Sakaguchi, Kyoko Shinzawa-Itoh, Shinya Yoshikawa, Hiroshi Fujii, Takashi Ogura
  • Organizer: 第47回生物物理学会年会
  • Place of Presentation: 徳島
• [Presentation] Structural Relaxation Process of Cytochrome c Oxidase Following Photolysis of Carbonmonoxide (2009)
  • Author(s): Izumi Ishigami, Satoru Nakashima, Kyoko Shinzawa-Itoh, Shinya Yoshikawa, Takashi Ogura
  • Organizer: 第47回生物物理学会年会
  • Place of Presentation: 徳島
• [Presentation] Structural analysis of FMN and iron-sulfer clusters of the bovine heart NADH-ubiquinone oxidoreductase by resonance Raman spectra (2009)
  • Author(s): Masahide Hikita, Masakazu Moriyama, Takashi Ogura, Kyoko Shinzawa-Itoh, Shinya Yoshikawa
  • Organizer: 第47回生物物理学会年会
  • Place of Presentation: 徳島
• [Presentation] Non-Resonance Raman Investigations of Bovine Mitochondrial F_o F_1-ATPsynthase (2009)
  • Author(s): Kaoru Mieda, Minoru Kubo, Kyoko Shinzawa-Itoh,Takashi Ogura, Shinya Yoshikawa
  • Organizer: 第47回生物物理学会年会
  • Place of Presentation: 徳島
• [Presentation] Detection of Two Oxygen-Isotope Sensitive Resonance Raman Bands for Human Indoleamine 2, 3-Dioxygenase (2009)
  • Author(s): Sachiko Yanagisawa, Keiko Yotsuya, Yumi Hashiwaki, Masaki Horitani, Hiroshi Sugimoto, Yoshitsugu Shiro, Takashi Ogura
  • Organizer: 第47回生物物理学会年会
  • Place of Presentation: 徳島
• [Presentation] Protein Dynamics of Cytochrome c Oxidase as Studied with Resonance Raman Spectroscopy (2009)
  • Author(s): Takashi Ogura, Satoru Nakashima, Izumi Ishigami, Kyoko Shinzawa-Itoh, Shinya Yoshikawa
  • Organizer: Symposium on Advanced Biological Inorganic Chemistry (招待講演)
  • Place of Presentation: Mumbai, India
• [Presentation] チトクロム酸化酵素のP反応中間体に現れる酸素同位体敏感ラマン線(356cm^<-1>)の帰属決定 (2009)
  • Author(s): 小倉尚志, その他
  • Organizer: 日本化学会第89春季年会
  • Place of Presentation: 船橋市
• [Presentation] フェリルオキソ型ヘムタンパク質の赤色励起共鳴ラマン分光 (2009)
  • Author(s): 池村賢一郎, その他
  • Organizer: 日本化学会第89春季年会
  • Place of Presentation: 船橋市
• [Presentation] 時間分解共鳴ラマン分光法を用いたチトクロムc酸化酵素のCO光乖離後の構造緩和過程の追跡 (2009)
  • Author(s): 石上泉, その他
  • Organizer: 日本化学会第89春季年会
  • Place of Presentation: 船橋市
• [Presentation] ウシ心筋チトクロムC酸化酵素のプロトンポンプ反応 (2008)
  • Author(s): 小倉尚志
  • Organizer: 特定領域研究「高次系分子科学」ミニ公開シンポジウム
  • Place of Presentation: 豊中市
  • Year and Date: 2008-08-06
• [Presentation] 時間分解共鳴ラマン分光法を用いたチトクロムc酸化酵素のCO光乖離後の構造緩和過程の追跡 (2008)
  • Author(s): 石上泉その, 他
  • Organizer: 第35回生体分子科学討論会
  • Place of Presentation: 兵庫県赤穂郡
• [Presentation] Resonance Raman Spectroscopy of Cytochrome c Oxidase (2008)
  • Author(s): Takashi Ogura, et.al.
  • Organizer: The 4^<th> Asian Biological Inorganic Chemistry Conference
  • Place of Presentation: Jeju, Korea
• [Presentation] Assignment of the Oxygen-Isotope-Sensitive Resonance Raman Band at 356-cm^<-1> of the P-Intermediate of Cytochrome c Oxidase to an O=Fe-N Bending Mode (2008)
  • Author(s): Kenichiro Ikemura, et.al.
  • Organizer: The 4^<th> Asian Biological Inorganic Chemistry Conference
  • Place of Presentation: Jeju, Korea
• [Remarks]
  • URL: http://www.sci.u-hyogo.ac.jp/life/biophys2/MolBiophys_II_main/home.html