光駆動イオン輸送蛋白質の動作機構の解明

• Project Area: Innovative nanoscience of supermolecular motor proteins working in biomembranes
• Project/Area Number: 21023014
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Institute for Molecular Science
• Principal Investigator: 古谷 祐詞 分子科学研究所, 生命・錯体分子科学研究領域, 准教授 (80432285)
• Project Period (FY): 2009 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥6,200,000 (Direct Cost: ¥6,200,000); Fiscal Year 2010: ¥3,100,000 (Direct Cost: ¥3,100,000); Fiscal Year 2009: ¥3,100,000 (Direct Cost: ¥3,100,000)
• Keywords: ナノバイオ / 赤外分光法 / 生体分子 / 蛋白質 / 生物物理

Research Abstract

イオンポンプ蛋白質は、エネルギー生産や情報伝達に必要な膜電位やイオン濃度勾配の形成に重要な役割を果たしている。本研究では、高精度な赤外分光計測を駆使して、イオンが輸送される過程で生じるタンパク質の構造変化を明らかにし、その輸送機構の解明を行うことを目的としている。特に、イオンがタンパク質内部でどのような包摂状態にあるのかを、周辺に存在するアミノ酸側鎖や蛋白質内部の水分子の分子振動、さらには赤外活性なイオン分子(硝酸イオン)を用い、イオン分子そのものの分子振動から明らかにする。対象となるイオンポンプ蛋白質は光で駆動される古細菌型ロドプシンが主であるが、それらの研究を元に光駆動ではないイオンポンプ蛋白質(Na^+ポンプであるV-ATPaseなど)も研究対象とし、両者の駆動メカニズムの違いを明らかにする。
平成22年度は以下のような成果を得た。1)プロトンポンプ活性を示す古細菌型ロドプシンであるGloeobacter Rhodopsin(GR)に対して低温赤外分光計測を行い、光誘起構造変化および内部結合水を含む蛋白質内部の水素結合ネットワークの変化を明らかにした。その結果、古細菌型ロドプシンのプロトンポンプ活性と正の相関のある2400cm^<-1>以下の強い水素結合を形成した水分子がGRにも存在することがわかった(Hashimoto, et al.Biochemistry)。2)Na^+ポンプであるV-ATPaseのKリングにおけるイオン脱着に伴う構造変化を全反射赤外分光法によって明らかにした。分子量70万にもおよぶ膜タンパク質複合体においても酵素活性を保ったまま計測することが可能であることを示した(Furutani, et al.J.Am.Chem.Soc.)。

Research Products

• [Journal Article] Sodium or Lithium Ion-Binding-Induced Structural Changes in the K-ring of V-ATPase from Enterococcus hirae Revealed by ATR-FTIR Spectroscopy (2011)
  • Author(s): Yuji Furutani, et al.
  • Journal Title: J.Am.Chem.Soc. Volume: 133 Pages: 2860-2863
  • Peer Reviewed
• [Journal Article] Low-Temperature FTIR Study of Gloeobacter Rhodopsin : Presence of Strongly Hydrogen-Bonded Water and Long-Range Structural Protein Perturbation upon Retinal Photoisomerization (2010)
  • Author(s): Kyohei Hashimoto, et al.
  • Journal Title: Biochemistry Volume: 49 Pages: 3343-3350
  • Peer Reviewed
• [Journal Article] Protein Fluctuations as the Possible Origin of the Thermal Activation of Rod Photoreceptors in the Dark (2010)
  • Author(s): Victor Lorenz Fonfria, et al.
  • Journal Title: J.Am.Chem.Soc. Volume: 132 Pages: 5693-5703
  • Peer Reviewed
• [Journal Article] An FTIR Study of the Photoreaction of Bovine Rhodopsin in the Presence of Hydroxylamine (2010)
  • Author(s): Kouta Katayama, et al.
  • Journal Title: J.Phys.Chem.B Volume: 114 Pages: 9039-9046
  • Peer Reviewed
• [Journal Article] Molecular insight into intrinsic heme distortion in ligand binding in hemoprotein (2010)
  • Author(s): Saburo Neya, et al.
  • Journal Title: Biochemistry Volume: 49 Pages: 5642-5650
  • Peer Reviewed
• [Journal Article] An FTIR Study of Monkey Green- and Red-Sensitive Visual Pigments (2010)
  • Author(s): Kouta Katayama et al.
  • Journal Title: Angew.Chem.Int.Ed. 49 Pages: 891-894
  • Peer Reviewed
• [Journal Article] Manipulation of the Protein-complex Function by Using an Engineered Heterotrimeric Coiled-coil Switch (2009)
  • Author(s): Toshihisa Mizuno et al.
  • Journal Title: Org.Biomol.Chem. 7 Pages: 3102-3111
  • Peer Reviewed
• [Journal Article] Effects of Chloride Ion Binding on the Photochemical Properties of Salinibacter Sensory Rhodopsin I (2009)
  • Author(s): Daisuke Suzuki et al.
  • Journal Title: J.Mol.Biol. 392 Pages: 48-62
  • Peer Reviewed
• [Journal Article] Engineering an inward proton transport from a bacterial sensor rhodopsin (2009)
  • Author(s): Akira Kawanabe et al.
  • Journal Title: J.Am.Chem.Soc. 131 Pages: 16439-16444
  • Peer Reviewed
• [Journal Article] Characterization of signaling complex composed of sensory rhodopsin I and its cognate transducer protein from the eubacterium Salinibacter rubber (2009)
  • Author(s): Yuki Sudo et al.
  • Journal Title: Biochemistry 48 Pages: 10136-10145
  • Peer Reviewed
• [Journal Article] Interaction between Na+ ion and carboxylates of the PomA-PomB stator unit studied by ATR-FTIR spectroscopy (2009)
  • Author(s): Yuki Sudo et al.
  • Journal Title: Biochemistry 48 Pages: 11699-11705
  • Peer Reviewed
• [Presentation] 赤外分光法でみるV型ATPaseのイオン結合・解離に伴う構造変化 (2011)
  • Author(s): 古谷祐詞
  • Organizer: 平成22年度日本生物物理学会中部支部討論会
  • Place of Presentation: 名古屋大学野依学術交流センター(愛知)
  • Year and Date: 2011-03-31
• [Presentation] Ion-binding-induced Structural Changes in Membrane Proteins Studied by ATR-FTIR Spectroscopy (2011)
  • Author(s): Yuji Furutani
  • Organizer: Third Korea-Japan Seminars on Biomolecular Sciences : Experiments and Simulations
  • Place of Presentation: Lotte Hotel, Jeju,(韓国)
  • Year and Date: 2011-02-27
• [Presentation] FTIR Studies of Membrane Proteins (2011)
  • Author(s): Yuji Furutani
  • Organizer: The Winter School of Sokendai/Asian CORE Program "Frontiers of Molecular Science-Life, Material, Energy, and Space"
  • Place of Presentation: 岡崎コンファレンスセンター(愛知)
  • Year and Date: 2011-02-21
• [Presentation] Modification of Gold Thin Film Suitable for Surface-enhanced Vibrational Spectroscopy on Membrane Proteins (2010)
  • Author(s): Hao Guo
  • Organizer: 4^<th> International Symposium on Nanomedicine (ISNM2010)
  • Place of Presentation: 岡崎コンファレンスセンター(愛知)
  • Year and Date: 2010-11-30
• [Presentation] 時間分解FTIR差スペクトル計測によるファラオニスハロロドプシンの陰イオン輸送過程での構造変化解析 (2010)
  • Author(s): 木村哲就
  • Organizer: 第36回日本生体エネルギー研究会&特定領域研究「革新的ナノバイオ」合同シンポジウム
  • Place of Presentation: 大阪大学銀杏会館(吹田)
  • Year and Date: 2010-11-19
• [Presentation] X-D伸縮振動領域を含めた時間分解赤外分光法によるバクテリオロドプシンの光誘起構造変化解析 (2010)
  • Author(s): 古谷祐詞
  • Organizer: 第36回日本生体エネルギー研究会&特定領域研究「革新的ナノバイオ」合同シンポジウム
  • Place of Presentation: 大阪大学銀杏会館(大阪)
  • Year and Date: 2010-11-18
• [Presentation] Modification of Gold Thin Film Suitable for Surface-enhanced Vibrational Spectroscopy on Membrane Proteins (2010)
  • Author(s): Hao Guo
  • Organizer: The Overseas Sokendai Lecture in Bangkok 2010
  • Place of Presentation: Chulalongkorn University, Bangkok(タイ)
  • Year and Date: 2010-10-20
• [Presentation] Stimulus-Induced Difference Infrared Spectroscopy for Membrane Proteins ; Visual and Archaeal-type Rhodopsins, and Flagella Motor Protein (2010)
  • Author(s): Yuji Furutani
  • Organizer: Indo-Japan Joint Workshop on "New Frontiers of Molecular Spectroscopy ; from Gas Phase to Proteins"
  • Place of Presentation: Hotel Kitano Plaza Rokkoso(兵庫)
  • Year and Date: 2010-09-28
• [Presentation] Time-resolved FT-IR measurement of X-D stretching vibrations of Bacteriorhodopsin in micro-to mill-second time range (2010)
  • Author(s): Yuji Furutani
  • Organizer: The 48th Annual Meeting of the Biophysical Society of Japan
  • Place of Presentation: 東北大学(宮城)
  • Year and Date: 2010-09-20
• [Presentation] Conformational Changes upon Chloride-ion Pumping During the pharaonis Halorhodopsin Photocycle Studied by Time-resolved FTIR Spectroscopy (2010)
  • Author(s): Testunari Kimura
  • Organizer: The 48th Annual Meeting of the Biophysical Society of Japan
  • Place of Presentation: 東北大学(仙台)
  • Year and Date: 2010-09-20
• [Presentation] 時間分解赤外分光法によるバクテリオロドプシンのX-D伸縮振動の解析 (2010)
  • Author(s): 古谷祐詞
  • Organizer: 第4回分子科学討論会
  • Place of Presentation: 大阪大学(大阪)
  • Year and Date: 2010-09-15
• [Presentation] 赤外分光法によるイオン輸送蛋白質の構造変化解析 (2010)
  • Author(s): 古谷祐詞
  • Organizer: アドバンス生命理学特論:世話人;須藤雄気准教授(名大・院理)
  • Place of Presentation: 名古屋大学(愛知)
  • Year and Date: 2010-07-13
• [Presentation] 光駆動イオン輸送蛋白質の動作機構の解明 (2010)
  • Author(s): 古谷祐詞
  • Organizer: 特定領域研究「革新的ナノバイオ」第5回班会議
  • Place of Presentation: 学習院大学(東京)
  • Year and Date: 2010-06-24
• [Presentation] Interaction between Membrane Proteins and Ions Studied by ATR-FTIR Spectroscopy : A Potential Tool for Drug Screening (2010)
  • Author(s): Yuji Furutani
  • Organizer: BIT's 1^<st> Annual International Conference of Medichem-2010
  • Place of Presentation: Beijing International Convention Center, Beijing,(中国)
  • Year and Date: 2010-05-19
• [Presentation] X-H, X-D伸縮振動領域の赤外分光法から何が分かるか? (2010)
  • Author(s): 古谷祐詞
  • Organizer: 分子研研究会「拡がるロドプシンの仲間から"何が分かるか""何をもたらすか"」
  • Place of Presentation: 岡崎コンファレンスセンター、岡崎
• [Presentation] 霊長類色覚視物質の赤外分光解析 (2010)
  • Author(s): 片山耕大
  • Organizer: 新学術領域「π空間」第3回公開シンポジウム
  • Place of Presentation: 岡崎コンファレンスセンター、岡崎
• [Presentation] KcsAチャネルの全反射赤外分光解析 (2010)
  • Author(s): 浅井佑介
  • Organizer: 新学術領域「π空間」第3回公開シンポジウム
  • Place of Presentation: 岡崎コンファレンスセンター、岡崎
• [Presentation] センサーロドプシンを用いたプロトンポンプ研究 (2010)
  • Author(s): 川鍋陽
  • Organizer: 分子研研究会「拡がるロドプシンの仲間から"何が分かるか""何をもたらすか"」
  • Place of Presentation: 岡崎コンファレンスセンター、岡崎
• [Presentation] 霊長類色覚視物質の赤外分光解析 (2010)
  • Author(s): 片山耕大
  • Organizer: 分子研研究会「拡がるロドプシンの仲間から"何が分かるか""何をもたらすか"」
  • Place of Presentation: 岡崎コンファレンスセンター、岡崎
• [Presentation] 高速原子間力顕微鏡によるロドプシンの動態観察―ロドプシンの直接観察から何が分かるか、何をもたらすか― (2010)
  • Author(s): 山下隼人
  • Organizer: 分子研研究会「拡がるロドプシンの仲間から"何が分かるか""何をもたらすか"」
  • Place of Presentation: 岡崎コンファレンスセンター、岡崎
• [Presentation] 霊長類色覚視物質の赤外分光解析 (2010)
  • Author(s): 片山耕大
  • Organizer: 平成21年度生物物理学会中部支部講演会
  • Place of Presentation: 岡崎コンファレンスセンター、岡崎
• [Presentation] ATR-FTIR Spectroscopy for Investigating Interaction Changes upon Binding of Ions to Transmembrane Proteins (2009)
  • Author(s): 古谷祐詞
  • Organizer: First World Congress of International Academy of Nanomedicine(IANM)
  • Place of Presentation: Sanya, China
• [Presentation] 赤外分光法による膜タンパク質の機能発現に伴う構造変化解析 (2009)
  • Author(s): 古谷祐詞
  • Organizer: 第49回分子科学若手の会夏の学校
  • Place of Presentation: もみのき森林公園、広島県廿日市市
• [Presentation] KcsAチャネルに対する赤外分光の狙いは達成されたのか? (2009)
  • Author(s): 古谷祐詞
  • Organizer: 特定領域研究「高次系分子科学」第7回ミニ公開シンポジウム
  • Place of Presentation: 越前町生涯学習センター越前分館、越前
• [Presentation] Interaction between membrane protein and ions characterized by ATR-FTIR spectroscopy (2009)
  • Author(s): 古谷祐詞
  • Organizer: The 47th Annual Meeting of the Biophysical Society of Japan
  • Place of Presentation: ASTY Tokushima, Tokushima
• [Presentation] Stimulus-Induced Difference FTIR Spectroscopy for Archaeal-Type Rhodopsins (2009)
  • Author(s): 古谷祐詞
  • Organizer: 2nd Japan-Korea Seminar on Biomolecular Sciences-Experiments and Simulations
  • Place of Presentation: Symposion Hall, Nagoya University, Nagoya
• [Presentation] Analysis of Hydrogen-Bonding Dynamics of a Water Cluster Formed in an Isolated Nanospace (2009)
  • Author(s): 古谷祐詞
  • Organizer: The 3rd Symposium on "Molecular Science for Supra Functional Systems"
  • Place of Presentation: Tokyo Institute of Technology, O-okayama
• [Presentation] 光駆動イオン輸送蛋白質の動作機構の解明 (2009)
  • Author(s): 古谷祐詞
  • Organizer: 特定領域研究「革新的ナノバイオ」第4回班会議
  • Place of Presentation: 北海道大学、札幌
• [Presentation] Structural Analysis of Sodium Ion Binding Sites of V-ATPase from Enterococcus hirae by use of ATR-FTIR Spectroscopy (2009)
  • Author(s): 古谷祐詞
  • Organizer: Internal Symposium : Innovative Nanoscience of Supermolecular Motor Proteins Working in Biomembranes
  • Place of Presentation: Shirankaikan in Kyoto University, Kyoto
• [Presentation] 全反射赤外分光法によるKcsAチャネルのイオン選択およびpHセンシング機構の解析 (2009)
  • Author(s): 古谷祐詞
  • Organizer: 第3回分子科学討論会
  • Place of Presentation: 名古屋大学、名古屋
• [Presentation] Sodium ion binding sites of V-ATPase studied by ATR-FTIR spectroscopy (2009)
  • Author(s): 古谷祐詞
  • Organizer: The 47th Annual Meeting of the Biophysical Society of Japan
  • Place of Presentation: ASTY Tokushima, Tokushima
• [Presentation] 古細菌型ロドプシンで実現した内向きのプロトンポンプ (2009)
  • Author(s): 川鍋陽
  • Organizer: 第36回生体分子科学討論会
  • Place of Presentation: 北海道大学学術交流会館、札幌
• [Presentation] 霊長類色覚視物質の赤外分光解析 (2009)
  • Author(s): 片山耕大
  • Organizer: 生物物理若手の会第49回夏の学校
  • Place of Presentation: 支笏湖ユースホステル、支笏湖
• [Presentation] ロドプシンのカーボンナノチューブへの内包 (2009)
  • Author(s): 川島崇睦
  • Organizer: 生物物理若手の会第49回夏の学校
  • Place of Presentation: 支笏湖ユースホステル、支笏湖
• [Presentation] ロドプシンのカーボンナノチューブへの内包 (2009)
  • Author(s): 川島崇睦
  • Organizer: 新学術領域研究「高次π空間」第2回公開シンポジウム
  • Place of Presentation: 京都テルサ、京都
• [Presentation] 霊長類色覚視物質の赤外分光解析 (2009)
  • Author(s): 片山耕大
  • Organizer: 第3回分子科学討論会
  • Place of Presentation: 名古屋大学、名古屋
• [Presentation] 古細菌型ロドプシンのカーボンナノチューブへの内包 (2009)
  • Author(s): 川島崇睦
  • Organizer: 第3回分子科学討論会
  • Place of Presentation: 名古屋大学、名古屋
• [Presentation] An FTIR Study of Monkey Green- and Red-Sensitive Visual Pigments (2009)
  • Author(s): 片山耕大
  • Organizer: Horiba-ISSP International Symposium(ISSP-11)on "Hydrogen and Water in Condensed Matter Physics"
  • Place of Presentation: Seimeinomori Resort, Chiba
• [Presentation] An FTIR Study of Monkey Green- and Red-Sensitive Visual Pigments (2009)
  • Author(s): 片山耕大
  • Organizer: The 47th Annual Meeting of the Biophysical Society of Japan
  • Place of Presentation: ASTY Tokushima, Tokushima
• [Presentation] ATR-FTIR study of KcsA channel (2009)
  • Author(s): 浅井佑介
  • Organizer: The 47th Annual Meeting of the Biophysical Society of Japan
  • Place of Presentation: ASTY Tokushima, Tokushima
• [Presentation] Study on chimeric archeal-rhodopsin containing cytoplasmic loop of bovine rhodopsin (2009)
  • Author(s): Aya Nakatsuma
  • Organizer: The 47th Annual Meeting of the Biophysical Society of Japan
  • Place of Presentation: ASTY Tokushima, Tokushima
• [Presentation] Encapsulation of ion-pumping rhodopsins into multi-wall carbon nanotubes (2009)
  • Author(s): 川島崇睦
  • Organizer: The 47th Annual Meeting of the Biophysical Society of Japan
  • Place of Presentation: ASTY Tokushima, Tokushima
• [Presentation] Study on chimeric archeal-rhodopsin containing cytoplasmic loop of bovine rhodopsin (2009)
  • Author(s): Aya Nakatsuma
  • Organizer: 1^<st> International Symposium on Emergence of Highly Elaborated π-Space and Its Function
  • Place of Presentation: Hotel Hankyu Expo Park, Suita