Motional linkage between voltage sensor and ion gate in voltage-dependent ion channels

• Project/Area Number: 21590040
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Physical pharmacy
• Research Institution: The University of Tokyo
• Principal Investigator: OSAWA Masanori 東京大学, 大学院・薬学系研究科, 助教 (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): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2011: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2010: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2009: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 構造生物学 / 電位依存性イオンチャネル / 電位センサードメイン / gating modifier toxin / 分子認識様式 / イオンチャネル / NMR / イオンゲート

Research Abstract

Voltage-dependent ion channels consist of a voltage-sensor domain(VSD) and a pore domain possessing the ion permeation pathway. This study demonstrated how VSD changes its conformation in a voltage-dependent manner, by a disulfide-locking method using a series of double cysteine mutants. In addition, the binding mode between VSD and its inhibiting toxin, analyzed by NMR, revealed the inhibition mechanism of the toxin.

Research Products

• [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] Backbone resonance assignments for Gproteinτ(i3) subunit in the GTP-boundstate (2012)
  • Author(s): Mase Y, Yokogawa M, Osawa M, Shimada I.
  • Journal Title: Biomol NMR Assign Volume: (in press)
  • Peer Reviewed
• [Journal Article] Backbone resonance assignments for the cytoplasmic region of the Mg^<2+> transporter MgtE in the Mg^<2+-> unboundstate (2012)
  • Author(s): Maruyama T, Imai S, Osawa M, Hattori M, Ishitani R, Nureki O, Shimada I.
  • Journal Title: Biomol NMR Assign Volume: (in press)
  • 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
• [Journal Article] NMR Analyses of the GβγBinding and Conformational Rearrangements of the Cytoplasmic Pore of G Protein-activated Inwardly Rectifying Potassium Channel 1(GIRK1) (2011)
  • Author(s): Yokogawa M, Osawa M, Takeuchi K, MaseY, Shimada I.
  • Journal Title: J. Biol. Chem Volume: 286 Issue: 3 Pages: 2215-2223
  • DOI: 10.1074/jbc.m110.160754
  • Peer Reviewed
• [Journal Article] Structure determination of a protein assembly by amino acid selective cross-saturation (2011)
  • Author(s): Kanamori E., Igarashi S., Osawa M., Fukunishi Y., Shimada I., Nakamura H.
  • Journal Title: Proteins Volume: 79 Issue: 1 Pages: 179-190
  • DOI: 10.1002/prot.22871
  • Peer Reviewed
• [Journal Article] NMR analyses of the Gβγ binding and conformational rearrangements of the cytoplasmic pore of G protein-activated inwardly rectifying potassium channel 1 (GIRK1). (2011)
  • Author(s): Yokogawa M.
  • Journal Title: J. Biol. Chem. Volume: 286 Pages: 2215-2223
  • Peer Reviewed
• [Journal Article] Quantitative characterization of Tob interactions provides the thermodynamic basis for translation termination-coupled deadenylase regulation (2010)
  • Author(s): Ruan L., Osawa M., Hosoda N., Imai S., Machiyama A., Katada T., Hoshino S., Shimada I.
  • Journal Title: J. Biol. Chem Volume: 285(36) Issue: 36 Pages: 27624-27631
  • DOI: 10.1074/jbc.m110.138867
  • Peer Reviewed
• [Journal Article] NMR analyses of the interaction between CCR5 and its ligand using functional reconstitution of CCR5 in lipid bilayers (2010)
  • Author(s): Yoshiura C, Kofuku Y, Ueda T, Mase Y, Yokogawa M, Osawa M, Terashima Y, Matsushima K, Shimada I
  • Journal Title: J Am Chem Soc. Volume: 132(19) Issue: 19 Pages: 6768-77
  • DOI: 10.1021/ja100830f
  • Peer Reviewed
• [Journal Article] Structural basis underlying the dual gate properties of KcsA (2010)
  • Author(s): Imai S, Osawa M, Takeuchi K, Shimada I
  • Journal Title: Proc Natl Acad Sci U S A Volume: 107(14) Issue: 14 Pages: 6216-21
  • DOI: 10.1073/pnas.0911270107
• [Journal Article] Backbone resonance assignments for the ligand binding subunit of the histidine permeasecomplex(HisJ) from Escherichia coli, under histidine-bound and unbound states (2010)
  • Author(s): Igarashi S., Osawa M., Ozawa S., Shimada I.
  • Journal Title: Biomol. NMR Assign Volume: 4(1) Issue: 1 Pages: 17-20
  • DOI: 10.1007/s12104-009-9200-6
  • Peer Reviewed
• [Journal Article] Quantitative characterization of Tob interactions provides the thermody namic basis for translation termination-counled deadenvlase regulation (2010)
  • Author(s): Ruan L.
  • Journal Title: J. Biol. Chem. Volume: 285 Pages: 27624-27631
  • Peer Reviewed
• [Journal Article] NMR analyses of the interaction between CCR5 and its ligand using functional reconstitution of CCR5 in lipid bilayers (2010)
  • Author(s): Yoshiura C.
  • Journal Title: J. Am. Chem. Soc. Volume: 132 Pages: 6768-6777
  • Peer Reviewed
• [Journal Article] Structural basis underlying the dual gate properties of KcsA (2010)
  • Author(s): Imai, S.
  • Journal Title: Proc.Natl.Acad.Sci.USA. 107 Pages: 6216-6221
  • Peer Reviewed
• [Journal Article] Backbone resonance assignments for the cytoplasmic regions of Gprotein-activated inwardly rectifying potassium channel 1(GIRK1) (2009)
  • Author(s): Yokogawa M., Muramatsu T., Takeuchi K., Osawa M., Shimada I.
  • Journal Title: Biomol. NMR Assign Volume: 3(1) Issue: 1 Pages: 125-128
  • DOI: 10.1007/s12104-009-9156-6
• [Journal Article] Evidence for the direct interaction of spermine with the inwardly rectifying potassium channel (2009)
  • Author(s): Osawa M., Yokogawa M., Muramatsu T., Kimura T., Mase Y., Shimada I.
  • Journal Title: J. Biol. Chem Volume: 284(38) Issue: 38 Pages: 26117-26126
  • DOI: 10.1074/jbc.m109.029355
  • Peer Reviewed
• [Journal Article] Cross-saturation and transferred cross-saturation experiments (2009)
  • Author(s): Shimada I., Ueda T., Matsumoto M., Sakakura M., Osawa M., Takeuchi K., Nishida N., Takahashi H.
  • Journal Title: Prog. Nuc. Magn. Reson. Spect Volume: 54 Issue: 2 Pages: 123-140
  • DOI: 10.1016/j.pnmrs.2008.07.001
  • Peer Reviewed
• [Journal Article] Evidence for the direct interaction of spermine with the inwardly rectifying potassium channel. (2009)
  • Author(s): Osawa M
  • Journal Title: J.Biol.Chem. 284 Pages: 26117-26126
  • Peer Reviewed
• [Journal Article] Backbone resonance assignments for the ligand binding subunit of the histidine permease complex(HisJ)from Escherichia coli, under histidine-bound and unbound states (2009)
  • Author(s): Igarashi S
  • Journal Title: Biomol.NMR Assign. (In press)
  • Peer Reviewed
• [Journal Article] Backbone resonance assignments for the cytoplasmic regions of G protein-activated inwardly rectifying potassium channel 1(GIRK1) (2009)
  • Author(s): Yokogawa M
  • Journal Title: Biomol.NMR Assign. 3 Pages: 125-128
  • Peer Reviewed
• [Journal Article] Cross-saturation and transferred cross-saturation experiments. (2009)
  • Author(s): Shimada I
  • Journal Title: Prog.Nuc.Magn.Reson.Spect. 54 Pages: 123-140
  • Peer Reviewed
• [Presentation] Structural basis for the inhibition of voltage-dependent potassium(Kv) channels by gating modifier toxins (2012)
  • Author(s): 小澤新一郎
  • Organizer: 第34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)
  • Year and Date: 2012-12-14
• [Presentation] Structural investigation of voltage-sensing mechanism of voltage-dependent K+channels (2011)
  • Author(s): 原田瞳
  • Organizer: 第34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)
  • Year and Date: 2011-12-14
• [Presentation] Structural investigation of voltage-sensing mechanism of voltage-dependent K^+ channels (2011)
  • Author(s): 原田瞳, 大澤匡範, 小澤新一郎, 木村友美, 野崎智裕, 嶋田一夫
  • Organizer: 第34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)
  • Year and Date: 2011-12-14
• [Presentation] Structural basis for the inhibition of voltage-dependent potassium (Kv) channels by gating modifier toxins (2011)
  • Author(s): 小澤新一郎, 大澤匡範, 木村友美, 原田瞳, 野崎智裕, 嶋田一夫
  • Organizer: 第34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)
  • Year and Date: 2011-12-14
• [Presentation] NMR analysis of the gating mechanism of inwardly rectifying K+channel, KirBac (2011)
  • Author(s): Toyama Y.
  • Organizer: 日本薬学会年会
  • Place of Presentation: ツインメッセ静岡(静岡)
  • Year and Date: 2011-03-29
• [Presentation] NMR analysis of the gating mechanism of inwardly rectifying K+ charmel, KirBac (2011)
  • Author(s): Toyama Y.
  • Organizer: 日本薬学会年会
  • Place of Presentation: ツインメッセ静岡(静岡)
  • Year and Date: 2011-03-29
• [Presentation] Structural basis for the specificity and efficiency of the G-protein signaling in GIRK-gating (2010)
  • Author(s): Mase Y.
  • Organizer: 日本分子生物学会・日本生化学会合同学会
  • Place of Presentation: 神戸ポートアイランド(兵庫)
  • Year and Date: 2010-12-10
• [Presentation] Development of residue selective cross-saturation method for modeling a large protein-protein complex (2010)
  • Author(s): Ozawa S.
  • Organizer: NMR討論会
  • Place of Presentation: タワーホール船堀(東京)
  • Year and Date: 2010-11-16
• [Presentation] Structural basis underlying the dual gate properties of KcsA (2010)
  • Author(s): Imai S.
  • Organizer: NMR討論会
  • Place of Presentation: タワーホール船堀(東京)
  • Year and Date: 2010-11-16
• [Presentation] Transferred Cross-Saturation Method under Magic Angle Spinning for Structural Analysis of Protein-Protein Interaction at the Solid-Liquid Interface (2010)
  • Author(s): Toyonaga S.
  • Organizer: NMR討論会
  • Place of Presentation: タワーホール船堀(東京)
  • Year and Date: 2010-11-16
• [Presentation] Development of residue selective cross-saturation method for modeling a large protein-protein complex (2010)
  • Author(s): Ozawa S.
  • Organizer: 国際生体系磁気共鳴学会
  • Place of Presentation: Cairns(オーストラリア)
  • Year and Date: 2010-08-16
• [Presentation] Structural basis underlying the dual gate properties of KcsA (2010)
  • Author(s): Imai S.
  • Organizer: 国際生体系磁気共鳴学会
  • Place of Presentation: Cairns(オーストラリア)
  • Year and Date: 2010-08-16
• [Presentation] Transferred Cross-Saturation Method under Magic Angle Spinning for Structural Analysis of Protein. Protein Interaction at the Solid. Liquid Interface (2010)
  • Author(s): Toyonaga S.
  • Organizer: 国際生体系磁気共鳴学会
  • Place of Presentation: Cairns(オーストラリア)
  • Year and Date: 2010-08-16
• [Presentation] Structural basis underlying the dual gate properties of KcsA (2010)
  • Author(s): Imai S.
  • Organizer: 国際生体系磁気共鳴学会
  • Place of Presentation: Caims(オーストラリア)
  • Year and Date: 2010-08-16
• [Presentation] Transferred Cross-Saturation Method under Magic Angle Spinning for Structural Analysis of Protein-Protein Interaction at the Solid-Liquid Interface (2010)
  • Author(s): Toyonaga S.
  • Organizer: 国際生体系磁気共鳴学会
  • Place of Presentation: Cairns(オーストラリア)
  • Year and Date: 2010-08-16
• [Presentation] Structural basis on the specificity and efficiency of the G-protein signaling for the GIRK-gating (2009)
  • Author(s): Mase Y.
  • Organizer: 日本分子生物学会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2009-12-09
• [Presentation] Structural basis of the inhibitory mechanism of collagen-binding to vWF andτ2υ1 integlin mediated by leech product LAPP (2009)
  • Author(s): Izawa N.
  • Organizer: 日本分子生物学会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2009-12-09
• [Presentation] Structural basis of the inhibitory mechanism of collagen-binding to vWF and α2βlinteglin mediated by leech product LAPP (2009)
  • Author(s): Izawa N.
  • Organizer: 日本分子生物学会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2009-12-09
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