Solid-state NMR analysis of membrane protein complexes with 7-transmembrane helices

• Project/Area Number: 19370068
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biophysics
• Research Institution: Osaka University
• Principal Investigator: FUJIWARA Toshimichi Osaka University, 蛋白質研究所, 教授 (20242381)
• Project Period (FY): 2007 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥18,330,000 (Direct Cost: ¥14,100,000、Indirect Cost: ¥4,230,000); Fiscal Year 2009: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2008: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000); Fiscal Year 2007: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
• Keywords: 構造生物学 / 生物物理 / 蛋白質 / 原子・分子物理 / ナノバイオ / 生体分子 / 核磁気共鳴 / 膜蛋白質 / 蛋白質立体構造 / 固体NMR

Research Abstract

Solid-state NMR enables the structural determination of biological molecular complexes in unoriented solids. We verified the usefulness of magic-angle spinning solid-state NMR based on dipolar and J couplings for the structural analysis of ^<13>C uniformly labeled biomolecular complexes. We applied solid-state NMR to halorhodopshin, mastoparan-X and 159-redisue transducer protein pHtrII in lipid bilayers. Solid-state NMR provided the detailed structure of mastoparan-X with membranes by combining NMR constraints with replica exchange molecular dynamics simulation using implicit membrane environment. This method was applied to NMR analysis of larger membrane proteins. ^1H-^<13>C correlations of mobile segments of pHtrII were analyzed.

Research Products

• [Journal Article] 1H-Detected 1H-1H Correlation Spectroscopy of a Stereo-Array Isotope Labeled Amino Acid under Fast Magic-Angle Spinning (2010)
  • Author(s): Hiroki Takahashi, Masatsune Kainosho, Hideo Akutsu, Toshimichi Fujiwara
  • Journal Title: J. Magn. Reson 203 Pages: 253-256
  • Peer Reviewed
• [Journal Article] Dynamic Nuclear Polarization Experiments at 14. 1 T for Solid-State NMR (2010)
  • Author(s): Yoh Matsuki, Hiroki Takahashi, Keisuke Ueda, Toshitaka Idehara, Isamu Ogawa, Mitsuru Toda, Hideo Akutsu, Toshimichi Fujiwara
  • Journal Title: Phys. Chem. Chem. Phys DOI:10.1039/c002268c(in press)
  • Peer Reviewed
• [Journal Article] Continuously Frequency Tunable High Power Sub-THz Radiation Source-Gyrotron FU CW VI for 600 MHz DNP-NMR spectroscopy (2010)
  • Author(s): Toshitaka Idehara, Kosuke Kosuga, La Agusu, Ryusuke Ikeda, Isamu Ogawa, Teruo Saito, Yoh Matsuki, Keisuke Ueda, Toshimichi Fujiwara
  • Journal Title: J. Infrared Millim. Terahz. Waves DOI10.1007/s10762-010-9643-y(in press)
  • Peer Reviewed
• [Journal Article] ^1H-Detected ^1H-^1H Correlation Spectroscopy of a Stereo-Array Isotope Labeled Amino Acid under Fast Magic-Angle Spinning (2010)
  • Author(s): Hiroki Takahashi, Masatsune Kainosho, Hideo Akutsu, Toshimichi Fuiiwara
  • Journal Title: Journal of Magnetic Resonance 203 Pages: 253-256
  • Peer Reviewed
• [Journal Article] 固体nmr分光法による生体分子複合体の構造解析 (2009)
  • Author(s): 藤原敏道
  • Journal Title: 日本物理学会誌, 「実験技術」 64,(3月号) Pages: 179-186
  • NAID: 110007138740
  • Peer Reviewed
• [Journal Article] 固体nmr膜蛋白質複合体構造解析技術(融合発展する構造生物学とケミカルバイオロジーの最前線) (2009)
  • Author(s): 藤原敏道
  • Journal Title: 蛋白質核酸酵素 54(12) Pages: 1499-1505
• [Journal Article] 固体NMR膜蛋白質複合体構造解析技術 (2009)
  • Author(s): 藤原敏道
  • Journal Title: 蛋白質 核酸 酵素 54 Pages: 1499-1505
  • Peer Reviewed
• [Journal Article] Fluid Mechanical Matching of H+-ATPsynthase Subunit c Ring with Lipid Membranes Revealed by 2H Solid-State NMR (2008)
  • Author(s): Masatoshi Kobayashi, Andrey V. Struts, Toshimichi Fujiwara, Michael F. Brown, Hideo Akutsu
  • Journal Title: Biophys. J. 94 Pages: 4339-4347
  • Peer Reviewed
• [Journal Article] A Magic-Angle-Spinning NMR Method for 1H-1H Distance Measurement Using Coherent Polarization Transfer in 13C-Labeled Organic Solids (2008)
  • Author(s): Hiroki Takahashi, Hideo Akutsu, Toshimichi Fujiwara
  • Journal Title: J. Chem. Phys 129154504 Pages: 1-12
  • Peer Reviewed
• [Journal Article] H+-ATP合成酵素のエネルギー変換機構とそこにおけるソフトな相互作用(特集:ソフトな相互作用による膜インターフェースの機能制御) (2008)
  • Author(s): 阿久津秀雄, 八木宏昌, 藤原敏道
  • Journal Title: 生化学 第80巻,第10号 Pages: 907-916
• [Journal Article] A Magic-Angle-Spinning NMR Method for^1H-^1H Distance Measurement Using Coherent Polarization Transfer in ^<13>C-Labeled Organic Solids (2008)
  • Author(s): H. Takahashi, H. Akutsu, T. Fujiwara
  • Journal Title: Journal of Chemical Physics 129 Pages: 154504-154504
  • Peer Reviewed
• [Journal Article] Fluid Mechanical Matching of H^+-ATPsynthase Subunit c Ring with Lipid Membranes Revealed by ^2H Solid-State NMR (2008)
  • Author(s): M. Kobayashi, A. V. Struts, T. Fujiwara, M. F. Brown, H. Akutsu
  • Journal Title: Biophysical Journal 94 Pages: 4339-4347
  • Peer Reviewed
• [Journal Article] H^+-ATP合成酵素のエネルギー変換機構とそこにおけるソフトな相互作用 (2008)
  • Author(s): 阿久津秀雄, 八木宏昌, 藤原敏道
  • Journal Title: 生化学 80 Pages: 907-916
  • NAID: 10022594584
• [Journal Article] Fluid Mechanical Matching of H^+-ATPsynthase Subunit c Ring with Lipid Membranes Revealed by 2H Solid-State NMR (2008)
  • Author(s): Masatoshi Kobayashi
  • Journal Title: Biophys. J. (印刷中)
  • Peer Reviewed
• [Journal Article] Structure of the Light-Harvesting Bacteriochlorophyll c Assembly in Chlorosomes from Chlorobium Limicola Determined by Solid-State NMR (2007)
  • Author(s): A. Egawa, T. Fujiwara, T. Mizoguchi, Y. Kakitani, Y. Koyama, H. Akutsu
  • Journal Title: Proc. Natl. Acad. Sci. U.S. A 104 Pages: 790-795
  • Peer Reviewed
• [Journal Article] Detailed Consideration of Experimental Results of Gyrotron FUCWII Developed as a Radiation Source for DNP-NMR Spectroscopy (2007)
  • Author(s): La Agusu, T. Idehara, I. Ogawa, T. Saito, T. Kanamaki, H. Takahashi., T. Fujiwara
  • Journal Title: Int. J. Infrared Millim. Waves 28 Pages: 499-511
  • Peer Reviewed
• [Journal Article] Spectral Fitting for Signal Assignment and Structural Analysis of Uniformly 13C-Labeled Solid Proteins by Simulated Annealing Based on Chemical Shifts and Spin Dynamics (2007)
  • Author(s): Y. Matsuki, H. Akutsu, T. Fujiwara
  • Journal Title: J. Biomol. NMR 38 Pages: 325-339
  • Peer Reviewed
• [Journal Article] Structural Analysis of Pituitary Adenylate Cyclase-Activating Polypeptides Bound to Phospholipid Membranes by Magic Angle Spinning Solid-State NMR (2007)
  • Author(s): Nobuyasu Komi, Kayo Okawa, Yukihiro Tateishi, Masahiro Shirakawa, Toshimichi Fujiwara, Hideo Akutsu
  • Journal Title: Biochimica et Biophysica Acta-Biomembranes 1768 Pages: 3001-3011
  • Peer Reviewed
• [Journal Article] Spectral Fitting for Signal Assignment and Structural Analysis of Uniformly 13C-Labeled Solid Proteins by Simulated Annealing Based on Chemical Shifts and Spin Dynamics (2007)
  • Author(s): Y. Matsuki
  • Journal Title: J. Biomol. NMR 38 Pages: 325-339
  • Peer Reviewed
• [Journal Article] Structural Analysis of Pituitary Adenylate Cyclase-Activating Polypeptides Bound to Phospholipid Membranes by Magic Angle Spinning Solid-State NMR (2007)
  • Author(s): Nobuyasu Komi
  • Journal Title: Biochimica et Biophysica Acta-Biomembranes 1768 Pages: 3001-3011
  • Peer Reviewed
• [Presentation] 膜タンパク質ハロロドプシンの多次元固体NMR法による構造解析 (2009)
  • Author(s): 樋口真理花, 江川文子, 田巻初, 神谷昌克, 相沢智康, 河野敬一, 藤原敏道, 出村誠
  • Organizer: 第48回NMR討論会
  • Place of Presentation: 福岡
  • Year and Date: 2009-11-10
• [Presentation] Implicit membrane modelを用いた分子動力学計算と固体NMRスペクトルシュミレーションによる膜結合ペプチド・マストパランXの構造解析(ポスター発表) (2009)
  • Author(s): 池田恵介, 亀田倫史, 原田英里砂, 阿久津秀雄, 藤原敏道
  • Organizer: 第48回NMR討論会
  • Place of Presentation: 福岡
  • Year and Date: 2009-11-10
• [Presentation] Implicit membrane modelを用いた分子動力学計算と固体NMRスペクトルシミュレーションによる膜結合ペプチド・マストパランXの構造解析 (2009)
  • Author(s): 池田恵介, 亀田倫史, 原田英里砂, 阿久津秀雄, 藤原敏道
  • Organizer: 第48NMR討論会
  • Place of Presentation: 福岡、九州大学医学部百年講堂
  • Year and Date: 2009-11-10
• [Presentation] Method for the dynamic structural analysis of membrane proteins based on NMR、口頭 (2009)
  • Author(s): 藤原敏道
  • Organizer: 第47回日本生物物理学会
  • Place of Presentation: 徳島
  • Year and Date: 2009-11-01
• [Presentation] Method for the dynamic structural analysis of membrane proteins based on NMR (2009)
  • Author(s): 藤原敏道
  • Organizer: 第47日本生物物理学会
  • Place of Presentation: 徳島、アスティ徳島
  • Year and Date: 2009-11-01
• [Presentation] 多次元固体NMR法による膜蛋白質pHtrIIの構造解析 (2009)
  • Author(s): 江川文子, 佐伯美和子, 林こころ, 児島長次郎, 阿久津秀雄, 藤原敏道
  • Organizer: 第47日本生物物理学会
  • Place of Presentation: 徳島、アスティ徳島
  • Year and Date: 2009-11-01
• [Presentation] Solid-State NMR Analysis of Membrane Protein and High-Field DNP (2009)
  • Author(s): 藤原敏道
  • Organizer: 3rd Asia-Pacific NMR Symposium
  • Place of Presentation: Ramada Plaza Hotel, Jeju, Korea
  • Year and Date: 2009-10-26
• [Presentation] Mainchain structural analysis of transmembrane halobacterial tranceducer pHtrIIby multi-dimensional MAS solid-state NMR (2009)
  • Author(s): A. Egawa, K. Hayashi, C. Kojima, H. Akutsu, T. Fujiwara
  • Organizer: The 3rd Asia-Pacific NMR Symposium
  • Place of Presentation: Jeju, Korea
  • Year and Date: 2009-10-25
• [Presentation] Structural analysis of biomolecular complexes by solid-state NMR (2009)
  • Author(s): 藤原敏道
  • Organizer: International Symposium on Biological NMR
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2009-10-23
• [Presentation] Solid-state NMR structural analysis of membrane proteins and sensitivity enhancement (2009)
  • Author(s): Toshimichi Fujiwara
  • Organizer: 2nd Symposium on Structural Analysis of Biological Macromolecules & 7th Japan-Korea Bilateral Symposium on Biological NMR
  • Place of Presentation: 札幌
  • Year and Date: 2009-07-27
• [Presentation] 固体nmr膜蛋白質複合体構造解析技術 (2008)
  • Author(s): 藤原敏道, 出村誠, 児嶋長次郎
  • Organizer: 第46回生物物理学会シンポジウム
  • Place of Presentation: 福岡
  • Year and Date: 2008-12-05
• [Presentation] 多次元固体NMR法による膜蛋白質pHtrIIの主鎖構造解析 (2008)
  • Author(s): 江川文子, 林こころ, 児嶋長次郎, 阿久津秀雄, 藤原敏道
  • Organizer: 日本生物物理学会
  • Place of Presentation: 福岡国際会議場
  • Year and Date: 2008-12-05
• [Presentation] 膜タンパク質固体NMR構造解析と高磁場DNP (2008)
  • Author(s): 藤原敏道, 高橋大樹, 江川文子, 佐伯美和子, 阿久津秀雄, 林こころ, 児島長次郎, 出原敏孝, 小川勇, 戸田充
  • Organizer: 第47回NMR討論会
  • Place of Presentation: 筑波
  • Year and Date: 2008-11-13
• [Presentation] 高磁場DNPと固体NMR生体分子複合体構造解析 (2008)
  • Author(s): 藤原敏道
  • Organizer: 大阪大学蛋白質研究所セミナー「先端磁気共鳴がもたらす生体系研究の新展開」
  • Place of Presentation: 大阪
  • Year and Date: 2008-07-29
• [Presentation] Structural analysis of transmembrane halobacterial tranceducer pHtrII by multi-dimensional high-resolution solid-state NMR (2008)
  • Author(s): 江川文子, 佐伯美和子, 林こころ, 児嶋長次郎, 阿久津秀雄, 藤原敏道
  • Organizer: 2nd Pacific Rim International Conference on Protein Science
  • Place of Presentation: ケアンズ
  • Year and Date: 2008-06-23
• [Presentation] Solid-State NMR for Structural Analysis of Biomolecular Complexes (2008)
  • Author(s): Toshimichi Fujiwara
  • Organizer: Korea-Japan Bilateral Symposium on Biological NMR
  • Place of Presentation: ソウル
  • Year and Date: 2008-04-25
• [Presentation] 13C-13C and 1H-1H Distance Analysis from the Polarization Transfer in Uniformly 13C-Labeled Organic Solids under Magic-Angle Spinning (2008)
  • Author(s): H. Takahashi, A. Egawa, H. Akutsu, T. Fujiwara
  • Organizer: the 49th Experimental NMR Conference, Asilomar Conference Grounds
  • Place of Presentation: Pacific Grove, CA, USA(口頭発表)
  • Year and Date: 2008-03-10
• [Presentation] 多次元固体NMR法による膜蛋白質pHtrIIの構造解析 (2007)
  • Author(s): 江川 文子
  • Organizer: 第45回日本生物物理学会年会
  • Place of Presentation: パシフィコ横浜会議センター
  • Year and Date: 2007-12-22
• [Presentation] Solid-state NMR of pharaonis halorhodopsin, Optimization of sample preparation (2007)
  • Author(s): Marika Higuchi, Ayako Egawa, Masakatsu Kamiya, Tomoyasu Aizawa, Keiichi Kawano, Toshimichi Fujiwara, Makoto Demura
  • Organizer: 第45回生物物理学会
  • Place of Presentation: パシフィコ横浜(口頭発表)
  • Year and Date: 2007-12-21
• [Book] タンパク質の同位体ラベルとnmr分光法 (2009)
  • Author(s): 藤原敏道, 池上貴久
  • Publisher: 化学同人
• [Book] タンパク質の同位体ラベルとNMR分光法 (2009)
  • Author(s): 長谷俊治・高尾敏文・高木淳一編、平野久、高尾敏文、豊田岐聡、有坂文雄、亀山啓一、池上貴久、藤原敏道、岩崎憲治、光岡薫、安永卓生
  • Total Pages: 244
  • Publisher: 化学同人
• [Book] Structural analysis of membrane systems by solid-state NMR (2008)
  • Author(s): Y. Todokoro, M. Kobayashi, T. Fujiwara, H. Akutsu
  • Publisher: Osaka University Press
• [Book] Chemistry, Physics, and Biology in Macromolecular Science (2008)
  • Author(s): Y. Todokoro, M. Kobayashi, T. Fujiwara, H. Akutsu
  • Publisher: Osaka University Press
• [Book] Secondary Structure Analysis of Proteins from Angle Dependent Interactions (2007)
  • Author(s): T. Fujiwara, H. Akutsu
  • Publisher: Springer
• [Book] “固体高分解能NMR"生命科学のための機器分析実験ハンドブック(実験医学別冊) (2007)
  • Author(s): 阿久津秀雄, 藤原敏道
  • Publisher: 羊土社
• [Remarks]
  • URL: http://www.protein.osaka-u.ac.jp/biophys/
• [Remarks]
  • URL: http://www.protein.osaka-u.ac.jp/biophys/
• [Remarks]
  • URL: http://www.protein.osaka-u.ac.jp/biophys/bussei.html