Development of Light-induced Membrane Proteins as a High-sensitiveMembrane Potential Sensor

2013 Fiscal Year Final Research Report

• Project/Area Number: 23657096
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biophysics
• Research Institution: Hokkaido University
• Principal Investigator: DEMURA Makoto 北海道大学, 大学院・先端生命科学研究院, 教授 (70188704)
• Project Period (FY): 2012 – 2013
• Keywords: ハロロドプシン / オプトジェネティクス / 光遺伝学 / 膜タンパク質 / 光生物学

Research Abstract

Human eye has a light-sensitive membrane protein. This sensor protein called rhodopsin that can identify the color of the light. Contrary, a rhodopsin-like proteins were found in some microorganisms, which have a light-driven ion transport (membrane potential) features different from the eyes. In this study, in order to improve the light-driven ion transport of microbial rhodopsin, the modified rhodopsins were prepared and visible absorption spectra in equilibrium and transient states, and the trimeric structure of modified rhodopsins were investigated.

Research Products

• [Journal Article] Thermodynamic Parameters of Anion Binding to Halorhodopsin from Natronomonas pharaonis by Isothermal Titration Calorimetry (2013)
  • Author(s): S. Hayashi, J. Tamogami, T. Kikukawa, H. Okamoto, K. Shimono, S. Miyauchi, M . Demura, T. Nara, N. Kamo
  • Journal Title: Biophysical Chemistry Volume: 172 Pages: 61-67
  • DOI: DOI:10.1016/j.bpc.2013.01.001
  • Peer Reviewed
• [Journal Article] Salt Bridge in the Conserved His-Asp Cluster in Gloeobacter Rhodopsin Contributes to Trimer Formation (2013)
  • Author(s): T. Tsukamoto,T. Kikukawa, T. Kurata, K.H. Jung, N.Kamo, M. D emura
  • Journal Title: FEBS Lett Volume: 587 Pages: 322-327
  • DOI: DOI:10.1016/j.febslet.2012.12.022
  • Peer Reviewed
• [Journal Article] Formation and Dissociation of pharaonis Halorhodopsin in Detergent System (2012)
  • Author(s): J. Tamogami, T. Kikukawa, T. Nara, K
  • Journal Title: Biophysical J Volume: 102 Pages: 2906-2915
  • DOI: DOI:10.1016/j.bpj.2012.05.008
  • Peer Reviewed
• [Journal Article] Influence of halide binding on the hydrogen bonding network in the active site of Salinibacter sensory rhodopsin I (2012)
  • Author(s): L. Reissig, T. Iwata, T. Kikukawa, M. Demura, N. Kamo, H. Kandori, Y. Sudo
  • Journal Title: Biochemistry Volume: 51 Pages: 8802-8813
  • DOI: DOI:10.1021/bi3009592
  • Peer Reviewed
• [Journal Article] Dynamics of Dangling Bonds of Water Molecules in pharaonis Halorhodopsin during Chloride Ion Transportation (2012)
  • Author(s): Y. Furutani, K. Fujiwara, T. Kimura, T. Kikukawa, M . Demura, H. Kandori
  • Journal Title: J.Phys.Chem.Lett Volume: 3 Pages: 2964-2969
  • DOI: DOI:10.1021/jz301287n
  • Peer Reviewed
• [Journal Article] Homo-trimerFormation and Dissociation of pharaonisHalorhodopsin in Detergent System (2012)
  • Author(s): T. Tsukamoto, T. Sasaki, K. J. Fujimoto,T. Kikukawa, M. Kamiya, T. Aizawa, K. Kawano, N. Kamo, M. Demura
  • Journal Title: Biophysical J Volume: 102 Pages: 2906-2915
  • DOI: DOI:10.1016/j.bpj.2012.05.008
  • Peer Reviewed
• [Journal Article] Photo-induced Bleaching of Sensory Rhodopsin II (Phoborhodopsin)from Halobacterium salinarum by Hydroxylamine: Identification of the Responsible Intermediates (2012)
  • Author(s): J. Tamogami, T. Kikukawa, Y. Ikeda, M . Demura, T. Nara, N. Kamo
  • Journal Title: Journal of Photochemistry and Photobiology B: Biology Volume: 106 Pages: 87-94
  • DOI: DOI:10.1016/j.jphotobiol.2011.10.009
  • Peer Reviewed
• [Journal Article] Expression of salinarum halorhodopsin in Escherichia coli cells: solubilization in the presence of retinal yields the natural state (2011)
  • Author(s): K. Muroda, K. Nakashima, M. Shibata,Y. Yamashita, T. Kikukawa, T. Tsukamoto, M. Kamiya, T. Aizawa, K. Kawano, S. Miyauchi, N. Kamo, M. Demura
  • Journal Title: Biochim. Biophys. Acta Volume: 1808 Pages: 2905-2912
  • DOI: DOI:10.1016/j.bbamem.2011.08.035(11)
  • Peer Reviewed
• [Journal Article] NMR analysis of the fibronectin cell-adhesive sequence, Arg-Gly-Asp, in a recombinant silk-like protein and a model peptide (2011)
  • Author(s): T. Asakura, H. Nishi, A. Nagano, A. Yoshida, Y. Nakazawa, M. Kamiya, M. Demura
  • Journal Title: Biomacromolecules Volume: 12 Pages: 3910-3916
  • DOI: DOI:10.1021/bm2011196
  • Peer Reviewed
• [Journal Article] An amino acid residue (S201) in the retinal binding pocket regulates the photoreaction pathway of phoborhodopsin (2011)
  • Author(s): G. Dai, Y. Zhang, J. Tamogami, M . Demura, N. Kamo, H. Kandori, T. Iwasa
  • Journal Title: Biochemistry Volume: 50 Pages: 7177-7183
  • DOI: DOI:10.1021/bi200598r
  • Peer Reviewed
• [Presentation] 固体NMRにおけるタンパク質連鎖帰属へのGFTNMR法の応用 (2012)
  • Author(s): 田巻初、江川文子、神谷昌克、菊川峰志、相沢智康、河野敬一、藤原敏道出村誠
  • Organizer: NMR討論会
  • Place of Presentation: ウィンクあいち(名古屋市)
  • Year and Date: 20121108-10
• [Presentation] Homo-trimeric structure of Gloeobacter rhodopsin is regulated by the protonation state of Asp121, a counterion of the protonated Schiff base (2012)
  • Author(s): Takashi Tsukamoto, Takashi Kikukawa,Masakatsu Kamiya, Tomoyasu Aizawa, Keiichi Kawano, Kwang-Hwan Jung, Naoki Kamo and Makoto Demura
  • Organizer: 15th Int Conf. Retinal Proteins
  • Place of Presentation: Monte Verita, Ascona, (Switzerland)
  • Year and Date: 20120930-1005
• [Presentation] Structure and Molecularhalorhodopsin Function of Halorhodopsin (2012)
  • Author(s): Demura Makoto
  • Organizer: 日本生物物理学会年会
  • Place of Presentation: 名古屋大学東山キャンパス(名古屋市)
  • Year and Date: 20120922-24
• [Presentation] Photo-induced proton transfer of Anabaena sensory rhodopsin (2012)
  • Author(s): 長谷見崇俊、菊川峰志、神谷昌克、相沢智康、河野敬一、Kwang-Hwan Jung、加茂直樹、出村誠
  • Organizer: 日本生物物理学会
  • Place of Presentation: 名古屋大学東山キャンパス(名古屋市)
  • Year and Date: 20120921-24
• [Presentation] Homo-trimeric assembly of cyanobacterial ion-pump Gloeobacterhodopsin グロエオバクターロドプシンの三量体形成に関する研究 (2012)
  • Author(s): 塚本卓、菊川峰志、神谷昌克、相沢智康、河野敬一、Jung Kwang-Hwan、加茂直樹、出村誠
  • Organizer: 日本生物物理学会
  • Place of Presentation: 名古屋大学東山キャンパス(名古屋市)
  • Year and Date: 20120921-24
• [Presentation] 三次元固体NMR法による膜タンパク質ハロロドプシンの研究 (2012)
  • Author(s): 田巻初、江川文子、神谷昌克、菊川峰志、相沢智康、河野敬一、藤原敏道,出村誠
  • Organizer: 日本蛋白質科学会年会
  • Place of Presentation: 名古屋国際会議場(名古屋市)
  • Year and Date: 20120620-22
• [Presentation] ハロロドプシンの光反応サイクル中に起こるカロテノイドの吸光度変化 (2012)
  • Author(s): 菊川峰志、小久保麻美、塚本卓、橋本革、井原邦夫、加茂直樹、出村誠
  • Organizer: 日本蛋白質科学会年会
  • Place of Presentation: 名古屋国際会議場(名古屋市)
  • Year and Date: 20120620-22
• [Presentation] Homo-trimerパク質ハロロドプシンの研究 (2012)
  • Author(s): T. Tsukamoto, T. Sasaki, K. J. Fujimoto, T. Kikukawa, M. Kamiya, T. Aizawa, K. Kawano, N. Kamo, M. Demura
  • Organizer: 日本蛋白質科学会年会
  • Place of Presentation: 名古屋国際会議場(名古屋市)
  • Year and Date: 20120620-22
• [Presentation] ThreeDimensional Solid-state NMR study of 7TM-Halorhodopsin, ISNMR2011 (2011)
  • Author(s): Hajime Tamaki, Marika Higuchi, Ayako Egawa, Masakatsu Kamiya, Takashi Kikukawa,Tomoyasu Aizawa, Keiichi Kawano,Toshimichi Fujiwara, Makoto Demura
  • Organizer: The 50th Annual Meeting of the NMR Society of Japan
  • Place of Presentation: 大さん橋ホール(横浜市)
  • Year and Date: 20111115-18
• [Presentation] 膜蛋白質・ハロロドプシンの第一細胞外ループペプチドの発現・精製とNMR解析 (2011)
  • Author(s): 高梨善彦、神谷昌克、熊木康裕、菊川峰志、相沢智康、河野敬一、出村誠
  • Organizer: ペプチド討論会
  • Place of Presentation: 札幌コンベンションセンター(札幌市)
  • Year and Date: 20110927-29
• [Presentation] Volume change during the Cl- transport of Natronomonas pharaonis halorhodopsin (2011)
  • Author(s): Kousuke Shibasaki, TakashiKikukawa, Masakatsu Kamiya, TomoyasuAizawa, Keichi Kawano, Naoki Kamo,Makoto Demura
  • Organizer: 日本生物物理学会年会
  • Place of Presentation: 兵庫県立大学姫路書写キャンパス(兵庫県)
  • Year and Date: 20110916-18
• [Book] オプ トジェネティクス(光遺伝学)-光工学と遺伝子による行動制御技術の最前線 (2013)
  • Author(s): 出村誠
  • Total Pages: 24-33
  • Publisher: エヌ・ティー・エス、ハロロドプシンの構造と分子機能解析
• [Book] 広がるNMRの世界-40人の研究者からの熱いメッセージ (2011)
  • Author(s): 出村誠
  • Total Pages: 42-45
  • Publisher: コロナ社
• [Remarks] タンパク質にかくされた"プロテイン・コード"を解く!
  • URL: http://www.lfsci.hokudai.ac.jp/labs/infMakotoDemuraWhendoeshalorhodopsinana/research2.html
• [Remarks] ダイナミクスインフォマティクス「光で動くタンパク質・イオンポンプ」
  • URL: http://www.lfsci.hokudai.ac.jp/labs/infana/topics/2006/index.html