Voltage-dependent structural changes and functinal mechanism of voltage-dependent ion channels

• Project/Area Number: 24590048
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Physical pharmacy
• Research Institution: The University of Tokyo
• Principal Investigator: OSAWA Masanori 東京大学, 薬学研究科(研究院), 講師 (60361606)
• Co-Investigator(Kenkyū-buntansha): SHIMADA Ichio 東京大学, 大学院薬学系研究科, 教授 (70196476)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 電位依存性イオンチャネル / SS-locking / 構造変化 / 電位依存性K+チャネル / 膜電位依存性 / NMR / rHDL

Outline of Final Research Achievements

Voltage-dependent K+ channels (Kv) regulates active potential by selectively permeating K+ through cell membrane, playing critical roles in neurotransmission and muscle contraction. To date, voltage-dependent confomational changes of Kv remais elusive, due to lack of the method to analyze the strucutre under membrane potential.
The aim of this study is to reveal structrural mechanism of Kv that senses membrane potential. We established a novel method to analyze structure under membrane potential. Based on the results obtained here, we propose that Kv undergoes structural equilibrium that shifts depending upon the membrane potential.

Research Products

• [Journal Article] Calmodulin and STIM proteins: Two major calcium sensors in the cytoplasm and endoplasmic reticulum (2015)
  • Author(s): Marshall CB, Nishikawa T, Osawa M, Stathopulos PB, Ikura M
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 印刷中 Issue: 1 Pages: 5-21
  • DOI: 10.1016/j.bbrc.2015.01.106
  • Peer Reviewed
• [Journal Article] Structural basis for the binding of the membrane-proximal C-terminal region of chemokine receptor CCR2 with the cytosolic regulator FROUNT (2014)
  • Author(s): Esaki K, Yoshinaga S, Tsuji T, Toda E, Terashima Y, Saitoh T, Kohda D, Kohno T, Osawa M, Ueda T, Shimada I, Matsushima K, Terasawa H
  • Journal Title: FEBS J Volume: 281 Issue: 24 Pages: 552-566
  • DOI: 10.1111/febs.13096
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Cross-saturation and transferred cross-saturation experiments (2014)
  • Author(s): Ueda T, Takeuchi K, Nishida N, Stampoulis P, Kofuku Y, Osawa M, Shimada I
  • Journal Title: Q Rev Biophys Volume: 47 Issue: 2 Pages: 143-187
  • DOI: 10.1017/s0033583514000043
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Functional dynamics of cell surface membrane proteins (2014)
  • Author(s): Nishida N, Osawa M, Takeuchi K, Imai S, Stampoulis P, Kofuku Y, Ueda T, Shimada I
  • Journal Title: J Magn Reson Volume: 241 Pages: 86-96
  • DOI: 10.1016/j.jmr.2013.11.007
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Backbone resonance assignments for G protein αi3 subunit in the GDP-bound state. (2014)
  • Author(s): Mase Y., Yokogawa M., Osawa M., and Shimada I.
  • Journal Title: Biomol. NMR Assign. Volume: 8 Issue: 2 Pages: 237-241
  • DOI: 10.1007/s12104-013-9491-5
  • Peer Reviewed
• [Journal Article] Solution NMR study on functional mechanism of membrane proteins (2013)
  • Author(s): Osawa M., Imai S., Takeuchi K, and Shimada I.
  • Journal Title: Seibutsubutsuri Volume: 53 Pages: 236-241
  • Peer Reviewed
• [Journal Article] Backbone resonance assignments for the cytoplasmic region of the Mg(2+) transporter MgtE in the Mg (2+)-unbound state (2013)
  • Author(s): Maruyama T, Imai S, Osawa M, Hattori M, Ishitani R, Nureki O, Shimada I.
  • Journal Title: Biomol NMR Assign Volume: 7 Issue: 1 Pages: 93-96
  • DOI: 10.1007/s12104-012-9385-y
  • Peer Reviewed
• [Journal Article] Functional equilibrium of the KcsA structure revealed by NMR (2012)
  • Author(s): Imai S, Osawa M, Mita K, Toyonaga S, Machiyama A, Ueda T, Takeuchi K, Oiki S, Shimada I.
  • Journal Title: J Biol Chem Volume: 287 Issue: 47 Pages: 39634-39641
  • DOI: 10.1074/jbc.m112.401265
  • Peer Reviewed
• [Journal Article] Biological role of the two overlapping poly(A)-binding protein interacting motifs 2 (PAM2) of eukaryotic releasing factor eRF3 in mRNA decay. (2012)
  • Author(s): Osawa M, Hosoda N, Nakanishi T, Uchida N, Kimura T, Imai S, Machiyama A, Katada T, Hoshino S, Shimada I.
  • Journal Title: RNA Volume: 18 Issue: 11 Pages: 1957-1967
  • DOI: 10.1261/rna.035311.112
  • Peer Reviewed
• [Journal Article] Functional dynamics of proteins revealed by solution NMR (2012)
  • Author(s): Osawa M, Takeuchi K, Ueda T, Nishida N, Shimada I.
  • Journal Title: Curr Opin Struct Biol Volume: 22 Issue: 5 Pages: 660-669
  • DOI: 10.1016/j.sbi.2012.08.007
  • Peer Reviewed
• [Journal Article] Spatial distribution of cytoplasmic domains of the Mg(2+)-transporter MgtE, in a solution lacking Mg(2+), revealed by paramagnetic relaxation enhancement (2012)
  • Author(s): Imai S, Maruyama T, Osawa M, Hattori M, Ishitani R, Nureki O, Shimada I.
  • Journal Title: Biochim Biophys Acta Volume: 1824 Issue: 10 Pages: 1129-1135
  • DOI: 10.1016/j.bbapap.2012.06.008
  • Peer Reviewed
• [Journal Article] Structural basis for the modulation of the gating property of G protein-gated inwardly rectifying potassium ion channel (GIRK)by the i/o-family G protein α subunit (Gdi_<i/o>) (2012)
  • Author(s): 間瀬瑶子
  • Journal Title: The Journal of Biological Chemistry Volume: 287 Issue: 23 Pages: 19537-19549
  • DOI: 10.1074/jbc.m112.353888
  • Peer Reviewed
• [Journal Article] Backbone resonance assignments for G protein α(i3) subunit in the GTP-bound state (2012)
  • Author(s): Mase Y, Yokogawa M, Osawa M, Shimada I.
  • Journal Title: Biomol NMR Assign Volume: 6 Issue: 2 Pages: 217-220
  • DOI: 10.1007/s12104-012-9361-6
  • Peer Reviewed
• [Presentation] Gタンパク質三量体によるGタンパク質共役型内向き整流性カリウムチャネル活性制御機構の構造生物学的解明 (2014)
  • Author(s): 加納 花穂, 外山 侑樹, 岩橋 優太, 間瀬 瑶子, 横川 真梨子, 大澤 匡範, 嶋田 一夫
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] メチルTROSY法による膜タンパク質の機能調節メカニズムの解明 (2014)
  • Author(s): 大澤匡範, 今井駿輔, 竹内 恒, 嶋田一夫
  • Organizer: NMR討論会
  • Place of Presentation: 大阪大学（大阪府吹田市）
  • Year and Date: 2014-11-04 – 2014-11-06
  • Invited
• [Presentation] Structural Basis for Regulation of G Protein-activated Inwardly Rectifying Potassium (2014)
  • Author(s): Osawa M, Mase Y, Yokogawa M,Takeuchi K, Shimada I
  • Organizer: 日本生生物物理学会年会
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市）
  • Year and Date: 2014-09-25 – 2014-09-27
  • Invited
• [Presentation] Structural Basis for Regulation of G Protein-activated Inwardly Rectifying Potassium Channel 1 (GIRK1) by G Proteins (2014)
  • Author(s): Osawa M, Mase Y, Yokogawa M,Takeuchi K, Shimada I
  • Organizer: International Conference on Magnetic Resonance in Biological Systems
  • Place of Presentation: Dallas （USA）
  • Year and Date: 2014-08-24 – 2014-08-29
  • Invited
• [Presentation] Structural analyses of the gating mechanism of inwardly rectifying potassium channel (2014)
  • Author(s): Toyama Y, Osawa M, Yokogawa M, Shimada I
  • Organizer: International Conference on Magnetic Resonance in Biological Systems
  • Place of Presentation: Dallas （USA）
  • Year and Date: 2014-08-24 – 2014-08-29
• [Presentation] 電位依存性カリウムイオンチャネルの膜電位感受機構の解明 (2014)
  • Author(s): 野﨑智裕、小澤新一郎、原田瞳、木村友美、大澤匡範、嶋田一夫
  • Organizer: 第12回次世代を担う若手のためのフィジカル・ファーマフォーラム
  • Place of Presentation: 箱根高原ホテル（神奈川県足柄下郡）
  • Year and Date: 2014-07-14 – 2014-07-15
• [Presentation] 電位依存性カリウムイオンチャネルの膜電位感受機構の解明 (2014)
  • Author(s): 野﨑智裕, 小澤新一郎, 原田瞳, 木村友美, 大澤匡範, 嶋田一夫
  • Organizer: 日本薬学会 第134年会
  • Place of Presentation: 熊本大学（熊本県熊本市）
• [Presentation] イオンチャネルの機能に関連した動的構造 (2013)
  • Author(s): 今井駿輔、大澤匡範、三田建一郎、豊永翔、町山麻子、上田卓見、竹内 恒、老木成稔、嶋田一夫
  • Organizer: 第35回生体膜と薬物の相互作用シンポジウム
  • Place of Presentation: 東京大学（東京都文京区）
• [Presentation] Structural analyses of the gating mechanism of inwardly rectifying potassium channel
  • Author(s): Y. Toyama, M. Osawa, M. Yokogawa, S. Toyanaga, A, Machiyama, I. Shimada
  • Organizer: Program for Young Researcher Symposium in Europe
  • Place of Presentation: Oxford (英国)
• [Presentation] Structural analyses of the gating mechanism of inwardly rectifying potassium channel
  • Author(s): Y. Toyama, M. Osawa, M. Yokogawa, S. Toyanaga, A, Machiyama, I. Shimada
  • Organizer: 第12回 日本蛋白質科学会年会
  • Place of Presentation: 名古屋国際会議場 （愛知）
• [Presentation] Structural basis for the modulation of the gating property of G protein-gated inwardly rectifying potassium ion channel (GIRK) by the i/o-family G protein α subunit (Gαi/o)
  • Author(s): Yoko Mase, Mariko Yokogawa, Masanori Osawa, and Ichio Shimada
  • Organizer: The XXVth International Conference on Magnetic Resonance in Biological Systems (ICMRBS)
  • Place of Presentation: Lyon （フランス）
• [Presentation] Structural Elucidation of Mg2+-dependent Gating Mechanism of a Mg2+ Transporter MgtE
  • Author(s): T. Maruyama, S. Imai, M. Osawa, M. Hattori, R. Ishitani, O. Nureki, I. Shimada
  • Organizer: The XXVth International Conference on Magnetic Resonance in Biological Systems
  • Place of Presentation: Lyon （フランス）
• [Presentation] ATP effect on Mg2+-dependent gating of a Mg2+ channel MgtE
  • Author(s): T. Maruyama, S. Imai, M. Osawa, M. Hattori, R. Ishitani, O. Nureki, I. Shimada
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡国際会議場(福岡)
• [Book] Large Protein Complexes Revealed by Solution-State NMR: G Proteins and G Protein-Activated Inwardly Rectifying Potassium Ion Channel (2013)
  • Author(s): Osawa M., Mase Y., Yokogawa M., Takeuchi K. and Shimada I.
  • Total Pages: 32
  • Publisher: Royal Society of Chemistry
• [Book] 【試料分析講座】「蛋白質の分析」, 分析化学会 (2012)
  • Author(s): 大澤匡範、今井駿輔、嶋田一夫.
  • Total Pages: 34
  • Publisher: 丸善
• [Remarks] 東京大学大学院薬学系研究科 生命物理化学教室
  • URL: http://ishimada.f.u-tokyo.ac.jp/public_html/paper/Paper_e.html
• [Remarks] 東京大学大学院薬学系研究科機能薬学専攻 生体分子機能学大講座 生命物理化学教室
  • URL: http://ishimada.f.u-tokyo.ac.jp/public_html/index_j.html