ミトコンドリアエネルギー変換を駆動する蛋白質複合体の機能の原子機構

2009 年度 研究成果報告書

• 研究課題/領域番号: 16087208
• 研究種目: 特定領域研究
• 配分区分: 補助金
• 審査区分: 生物系
• 研究機関: 兵庫県立大学
• 研究代表者: 吉川 信也 兵庫県立大学, 大学院・生命理学研究科, 教授 (40068119)
• 研究分担者: 村本 和優 兵庫県立大学, 大学院・生命理学研究科, 准教授 (50305679) ; 伊藤 恭子 兵庫県立大学, 大学院・生命理学研究科, 助教 (70206316)
• 研究期間 (年度): 2004 – 2009
• キーワード: 酸素反応機構 / 膜蛋白質 / ミトコンドリア / エネルギー変換 / X線結晶構造解 / チトクロム酸化酵素 / NADH-ユビキノン還元酵素

研究概要

チトクロム酸化酵素の全脂質の構造が決定され、His503がプロトンポンプと酸素還元の共役部位であること、休止型酵素の酸素還元部位には過酸化物が結合していること、Fe_<a3>^<2+>に結合しているO_2^-が1段階3電子還元されること、水経路がプロトンポンプの逆流を効率よく防ぐこと等が明らかになった。NADH-ユビキノン還元酵素とFoF1ATP合成酵素が二次元結晶化された。水溶液中の蛋白質に適用できる超高感度赤外分光装置が開発された。

研究成果

• [雑誌論文] Bovine cytochrome c oxidase otructures enable O2 reduction with minimization of active oxygen species and provide a proton pumping gate (2010)
  • 著者名/発表者名: K. Muramoto, K. Ohta, K. Shinzawa-Itoh, K. Kanda, M. Taniguchi, E. Yamashita, T. Tsukihara, S. Yoshikawa
  • 雑誌名: Proc. Natl. Acad. Sci. USA (in press)
  • 査読あり
• [雑誌論文] A Resonance Raman Band Assignable to the O7O Stretching Mode in the Resting Oxidized State of Bovine Heart Cytochrome c Oxidase (2010)
  • 著者名/発表者名: M. Sakaguchi, K. Shinzawa-Itoh, S. Yoshikawa, T. Ogura
  • 雑誌名: J. Bioenergetics and Biomembranes (in press)
  • 査読あり
• [雑誌論文] Cell-free synthesis of cytochrome c oxidase, a multicomponent membrane protein (2010)
  • 著者名/発表者名: Y. Katayama, K. Shimokata, M. Suematsu, T. Ogura, T. Tsukihara, S. Yoshikawa, H. Shimada
  • 雑誌名: J. Bioenergetics and Biomembranes (in press)
  • 査読あり
• [雑誌論文] X-ray structure of the NO-bound Cu(B) in bovine cytochrome c oxidase. (2010)
  • 著者名/発表者名: Ohta K, Muramoto K, Shinzawa-Itoh K, Yamashita E, Yoshikawa S, Tsukihara T
  • 雑誌名: Acta Cryst. F 66 ページ: 251-253
  • 査読あり
• [雑誌論文] Bovine heart NADH-ubiquinone oxidoreductae contains one molecule of ubiquinone with ten isoprenoide units as one of the cofactors. (2010)
  • 著者名/発表者名: K. Shinzawa-Itoh, J. Seiyama, H. Terada, R. Nakatsubo, K. Naoki, Y. Nakashima, S. Yoshikawa
  • 雑誌名: Biochemistry 49 ページ: 487-492
  • 査読あり
• [雑誌論文] A Peroxide Bridge between Fe and Cu Ions in the O2 Reduction Site of Fully Oxidized Cytochrome c Oxidase could suppress the Proton Pump (2009)
  • 著者名/発表者名: H. Aoyama, K. Muramoto, K. Shinzawa-Itoh, K. Hirata, E. Yamashita, T. Tsukihara, T. Ogura, S. Yoshikawa
  • 雑誌名: Proc. Natl. Acad. Sci. USA 106 ページ: 2165-2169
  • 査読あり
• [雑誌論文] Iron-sulfur cluster N5 is coordinated by a HXXXCXXCXXXXXC motife in the NuoG subunit of E. coli NADH:quinine oxidoreductase (Complex I) (2008)
  • 著者名/発表者名: Nakamura-Ogiso, E., Matsuno-Yagi, A., Yoshikawa, S., Yagi, T., Ohnishi, T.
  • 雑誌名: J. Biol. Chem. 283 ページ: 25979-25987
  • 査読あり
• [雑誌論文] Functional role of Coenzyme Q in the energy coupling of NADH-CoQ oxidoreductase (Complex I): Stabilization of the semiquinone state with application of inside-positive membrane potential to proteoliposomes (2008)
  • 著者名/発表者名: Ohnishi, To., Ohnishi, Ts., Shinzawa-Itoh, K., Yoshikawa, S.
  • 雑誌名: Biofctor 32 ページ: 13-22
  • 査読あり
• [雑誌論文] Red-Excitation Resonance Raman Analysis of the nFe=O Mode of Ferryl-Oxo Hemoproteins (2008)
  • 著者名/発表者名: Ikemura, K., Mukai, M., Shimada, H., Tsukihara, T., Yamaguchi, S., Shinzawa-Itoh, K., Yoshikawa, S., Ogura, T.
  • 雑誌名: J. Am. Chem. Soc. 130 ページ: 14384-14385
  • 査読あり
• [雑誌論文] Resonance Raman spectra of the FMN of the bovine heart NADH: coenzyme Q oxidoreductase, the largest membrane protein in the mitochondrial respiratory system (2007)
  • 著者名/発表者名: Sugiyama, H., Nakatsubo, R., Yamaguchi, S., Ogura, T., Shinzawa-Itoh, K. Yoshikawa, S.
  • 雑誌名: J. Bioenrg. Biomembr. 39 ページ: 145-148
  • 査読あり
• [雑誌論文] A histidine residue acting as a controlling site for dioxygen reduction and proton pumping by cytochrome c oxidase (2007)
  • 著者名/発表者名: Muramoto, K., Hirata, K., Shinzawa-Itoh, K., Yoko-o, S., Yamashita, E., Aoyama, H., Tsukihara, T., Yoshikawa, S.
  • 雑誌名: Proc. Natl. Acad. Sci. USA 104 ページ: 7881-7886
  • 査読あり
• [雑誌論文] The proton pumping pathway of bovine heart cytochrome c oxidase (2007)
  • 著者名/発表者名: K. Shimokata, Y. Katayama, H. Murayama, M. Suematsu, T. Tsukihara, K. Muramoto, H. Aoyama, S. Yoshikawa, H. Shimada
  • 雑誌名: Proc. Natl. Acad. Sci. USA 104 ページ: 4200-4205
  • 査読あり
• [雑誌論文] Structures and physiological roles of thirteen integral lipids of bovine heart cytochrome c oxidase (2007)
  • 著者名/発表者名: K. Shinzawa-Itoh, H. Aoyama, K. Muramoto, H. Terada, T. Kurauchi, Y. Tadehara, A. Yamasaki, T. Sugimura, S. Kurono, K. Tsujimoto, T. Mizushima, E. Yamashita, T. Tsukihara, S. Yoshikawa
  • 雑誌名: EMBO J. 26 ページ: 1713-1725
  • 査読あり
• [雑誌論文] A histidine residue acting as a controlling site for dioxygen reduction and proton pumping by cytochrome c oxidase (2007)
  • 著者名/発表者名: K. Muramoto, K. Hirata, K. Shinzawa-Itoh, S. Yoko-o, E. Yamashita, H. Aoyama, T. Tsukihara, S. Yoshikawa
  • 雑誌名: Proc. Natl. Acad. Sci. USA 104 ページ: 7881-7886
  • 査読あり
• [雑誌論文] Reaction mechanism of bovine heart cytochrome c oxidase (2006)
  • 著者名/発表者名: S. Yoshikawa, K. Muramoto, K. Shinzawa-Itoh, H. Aoyama, T, Tsukihara, T. Ogura, K. Shimokata, Y. Katayama, H. Shimnada
  • 雑誌名: Biochim. Biophys. Acta 1757 ページ: 395-400
  • 査読あり
• [雑誌論文] Proton pumping mechanism of bovine heart cytochrome c oxidase (2006)
  • 著者名/発表者名: S. Yoshikawa, K. Muramoto, K. Shinzawa-Itoh, H. Aoyama, T, Tsukihara, K. Shimokata, Y. Katayama, H. Shimnada
  • 雑誌名: Biochim. Biophys. Acta 1757 ページ: 1110-1116
  • 査読あり
• [雑誌論文] Absolute configuration of the hydroxyfarnesylethyl group of haem A, determined by X-ray structural analysis of bovine heart cytopchyrome c oxidase using methods applicable at 2. 8 A resolution (2005)
  • 著者名/発表者名: E. Yamashita(阪大), H. Aoyama(理研), M. Yao(理研), K. Muramoto, K. Shinzawa-Itoh, S. Yoshikawa, T. Tsukihara(阪大)
  • 雑誌名: Acta Cryst. D61 ページ: 1373-1377
  • 査読あり
• [雑誌論文] Reaction mechanism and phospholipids structures of bovine heart cytochrome c oxidase (2005)
  • 著者名/発表者名: S. Yoshikawa
  • 雑誌名: Biochem. Soc. Trans. ページ: 934-937
  • 査読あり
• [雑誌論文] Probing the oxygen activation reaction in intact whole mitochondria through analysis of molecular vibrations (2005)
  • 著者名/発表者名: T. Takahashi, S. Kuroiwa, T. Ogura, S. Yoshikawa
  • 雑誌名: J. Am. Chem.. Soc. 127 ページ: 9970-9971
  • 査読あり
• [雑誌論文] The intrinsic stability of the second intermediate following the dioxygen-bound form in the O2 reduction by cytochrome c oxidase (2004)
  • 著者名/発表者名: K. Oda, T. Ogura, E.H. Appelman, S. Yoshikawa
  • 雑誌名: FEBS Lett. 570 ページ: 161-165
  • 査読あり
• [学会発表] Bovine heart cytochrome c oxidase structures facilitaiting the four-electron redeuction of O2 in one-step and the effective proton pump. (2010)
  • 著者名/発表者名: S. Yoshikawa
  • 学会等名: Invited Seminar in Department of Physiology and Biophysics, Albert Einstein College of Medicine
  • 発表場所: New York, NY, USA
  • 年月日: 2010-01-07
• [学会発表] X-ray structures of CO, NO and Cyanide derivatives of bovine heart cytochrome c oxidase. (2009)
  • 著者名/発表者名: S. Yoshikawa
  • 学会等名: 54th National Meeting of the Italian Society of Biochemistry and Molecular Biology
  • 発表場所: Citta Universitatria, Catania Italy
  • 年月日: 20090923-20090927
• [学会発表] Structures of bovine cytochrome c oxidase facilitaiting the four-electron redeuction of O2 in one-step and the effective proton pump. (2009)
  • 著者名/発表者名: S. Yoshikawa
  • 学会等名: 14th International Conference on Biological Inorganic Chemistry
  • 発表場所: Nagoya Japan, Plenary Lecture
  • 年月日: 20090725-20090730
• [学会発表] Mutagenesis studies on the functional roles of possible proton transfer pathways in bovine heart cytochrome c oxidase (2009)
  • 著者名/発表者名: H. Shimada
  • 学会等名: 14th International Conference on Biological Inorganic Chemistry
  • 発表場所: Nagoya Japan
  • 年月日: 20090725-20090730
• [学会発表] Structure and dynamics of cytochrome c oxidase as studied with resonance Raman spectroscopy. (2009)
  • 著者名/発表者名: T. Ogura
  • 学会等名: 14th International Conference on Biological Inorganic Chemistry
  • 発表場所: Nagoya Japan
  • 年月日: 20090725-20090730
• [学会発表] Reaction mechanism of bovine heart cytochrome c oxidase (2008)
  • 著者名/発表者名: Yoshikawa, S.
  • 学会等名: 53rd National Meeting of Italian Society of Biochemistry and Molecular Biology
  • 発表場所: Riccione, Italy
  • 年月日: 20080923-20080926
• [学会発表] Evidence for H channel proton pump in bovine cytochrome c oxidase (2008)
  • 著者名/発表者名: Yoshikawa, S.
  • 学会等名: 15th European Bioenergetics Conference
  • 発表場所: Dublin, Ireland
  • 年月日: 20080719-20080724
• [学会発表] High resolution X-ray diffraction experiment of bovine cytochrome c oxidase (2008)
  • 著者名/発表者名: Suga M
  • 学会等名: The 15th European Bioenergetics Conference
  • 発表場所: Dublin, Ireland
  • 年月日: 20080719-20080724
• [学会発表] X-ray crystal structural analysis of cyanide binding cytochrome c oxidase (2008)
  • 著者名/発表者名: Mochizuki M
  • 学会等名: The 15th European Bioenergetics Conference
  • 発表場所: Dublin, Ireland
  • 年月日: 20080719-20080724
• [学会発表] A peroxide bridge between the two metals in the dinuclear center of the fully oxidized cytochrome c oxidase (2008)
  • 著者名/発表者名: Aoyama H
  • 学会等名: The 15th European Bioenergetics Conference
  • 発表場所: Dublin, Ireland
  • 年月日: 20080719-20080724
• [学会発表] X-ray structure of carbon monoxide at copper site of the dinuclear site of cytochrome c oxidase (2008)
  • 著者名/発表者名: Muramoto K
  • 学会等名: The 15th European Bioenergetics Conference
  • 発表場所: Dublin, Ireland
  • 年月日: 20080719-20080724
• [学会発表] Structural analyses for lipid/protein interactions in bovine heart cytochrome c oxidase (2008)
  • 著者名/発表者名: Shinzawa-Itoh K, Aoyama H, Muramoto K, Terada H, Kurauchi T, Tadehara Y, Yamasaki A, Sugimura T, Kurono S, Tsujimoto K, Mizushima T, Yamashita E, Tsukihara T, Yoshikawa S.
  • 学会等名: The 15th European Bioenergetics Conference
  • 発表場所: Dublin, Ireland
  • 年月日: 20080719-20080724
• [学会発表] X-ray structural analysis of zinc/cadmium inhibitory site in bovine heart cytochrome c oxidase (2008)
  • 著者名/発表者名: Muramoto K
  • 学会等名: The 15th European Bioenergetics Conference
  • 発表場所: Dublin, Ireland
  • 年月日: 20080719-20080724
• [学会発表] Synthesis of functional Paracoccus Denitrificans cytochrome c oxidase by Eschericia cali cell-free coupled transcription/translation system (2008)
  • 著者名/発表者名: Y. Katayama
  • 学会等名: The 15th European Bioenergetics Conference
  • 発表場所: Dublin, Ireland
  • 年月日: 20080719-20080724
• [学会発表] Mutations of possible proton-transfer pathways of bovine heart cytochrome c oxidase (2008)
  • 著者名/発表者名: K, Shimokata
  • 年月日: 20080719-20080724
• [学会発表] Proton Pumping Mechanism of Bovine Heart Cytochrome c Oxidase, Joint Meeting of the Biophysical Society (2008)
  • 著者名/発表者名: Yoshikawa, S.
  • 学会等名: 52nd Annual Meeting and 16th International Biophysics Congress
  • 発表場所: Long Beach, California, USA
  • 年月日: 20080202-20080206
• [学会発表] Lipid-Protein Interactions in Cytochrome c Oxidase、Keystone Symposia (2008)
  • 著者名/発表者名: Yoshikawa, S.
  • 学会等名: Molecular Basis for Biological Membrane Organization
  • 発表場所: Big Sky, Montana USA
  • 年月日: 20080112-20080118
• [学会発表] Reaction mechanism of bovine heart cytochrome c oxidase (2006)
  • 著者名/発表者名: S. Yoshikawa
  • 学会等名: The 14th European Bioenergetics Conference
  • 発表場所: Moscow Russia
  • 年月日: 20060722-20060727
• [学会発表] Proton pumping mechanism of bovine heart cytochrome c oxidase (2006)
  • 著者名/発表者名: S. Yoshikawa
  • 学会等名: Bioenergetics Electron transport systems in biomembranes organized by S. Yoshikawa and W.A. Cramer
  • 発表場所: Kyoto, Japan
  • 年月日: 20060618-20061823
• [学会発表] Mutational analyses of a hydrogen bond network in a bacterial cytochrome c oxidase which is homologous to that of the bovine heart enzyme (2006)
  • 著者名/発表者名: Y. Yukutake
  • 学会等名: 20th IUBMB International Congress of Biochemistry and Molecular Biology
  • 発表場所: Kyoto, Japan
  • 年月日: 20060618-20061823
• [学会発表] Experimental results indicating that heme a of bovine heart cytochrome c oxidase is the driving element of the proton-pumping (2005)
  • 著者名/発表者名: S. Yoshikawa
  • 学会等名: International Conference on Mitochondria, from Molecular Insight to Physiology and Pathology 40 Years of Bari Meetings
  • 発表場所: University of Bari, Bari Italy
  • 年月日: 20051217-20051222
• [学会発表] Mass spectral and X-ray structural analyses of lipids in bovine heart cytochrome c oxidase (2005)
  • 著者名/発表者名: S. Yoshikawa
  • 学会等名: BioScience 2005-from Gene to Systems
  • 発表場所: SECC, Glasgow, UK
  • 年月日: 20050717-20050721
• [学会発表] The low spin heme of cytochrome c oxidase as the driving element of the proton pumping process (2004)
  • 著者名/発表者名: S. Yoshikawa
  • 学会等名: 13th European Bioenergetics Conference, Colloquim ID, "Heme-Copper Oxygen Reductase"
  • 発表場所: Pisa, Italy
  • 年月日: 20040821-20040826
• [図書] Biophysical and Structural Aspects of Bioenrgetics(Marten Wikstrom ed. ) (2005)
  • 著者名/発表者名: S. Yoshikawa
  • 総ページ数: 55-71
  • 出版者: Royal Society of Chemistry
• [図書] Virtual Text,Ergito Cambridge MA USA(B. Lewin ed., Mitochondrial cytochrome coxidase in "Structures") (2005)
  • 著者名/発表者名: S. Yoshikawa