Proton Pumping ATPase and Roles of Acidic pH

• Project/Area Number: 20370048
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: Iwate Medical University
• Principal Investigator: FUTAI Masamitsu Iwate Medical University, 薬学部, 教授 (50012646)
• Co-Investigator(Kenkyū-buntansha): NAKANISHI Mayumi 岩手医科大学, 薬学部, 准教授 (20270506) ; GOTO Naomi 岩手医科大学, 薬学部, 助教 (80403971) ; SEKIYA Mizuki 岩手医科大学, 薬学部, 助教 (70509033)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥20,280,000 (Direct Cost: ¥15,600,000、Indirect Cost: ¥4,680,000); Fiscal Year 2010: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2009: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000); Fiscal Year 2008: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
• Keywords: V-ATPase / F-ATPase / 破骨細胞 / リソソーム / 阻害剤 / V型ATPase / F型ATPase / マクロファージ

Research Abstract

We have studied rotary mechanism of F-ATPase (ATP synthase) and V-ATPase (lysosome-type ATPase) under the physiological conditions. Significant results were obtained for biochemical and thermodynamic regulatory mechanism. We also studied roles of V-ATPase in vesicle trafficking and formation of osteoclast plasma membrane. Formation of multinuclear cells was studied together with osteoclast differentiation.

Research Products

• [Journal Article] Vacuolar type ATPases in animal and plant cells. (2011)
  • Author(s): H. Okamoto, M. Futai
  • Journal Title: Encyclopedia of Biophys. in press
• [Journal Article] The mechanism of rotating proton pumping ATPases. (2010)
  • Author(s): M. Nakanishi-Matsui, M. Sekiya, R.K. Nakamoto, M. Futai
  • Journal Title: Biochim.Biophys.Acta 1797 Pages: 1343-1352
• [Journal Article] Roles of the β subunit hinge domain in ATP synthase F_1 sector : Hydrophobic network formed by introduced β Phe174 inhibits subunit rotation. (2010)
  • Author(s): M. Nakanishi-Matsui, S. Kashiwagi, M. Kojima, T. Nonaka, M. Futai
  • Journal Title: Biochem.Biophys.Res.Commun. 395 Pages: 173-177
• [Journal Article] Single molecule behavior of inhibited and active states of Escherichia coli ATP synthase F_1 rotation. (2010)
  • Author(s): M. Sekiya, H. Hosokawa, M. Nakanishi Matsui, M.K. Al-Shawi, R.K. Nakamoto, M. Futai
  • Journal Title: J.Biol.Chem. 285 Pages: 42058-42067
• [Journal Article] The mechanism of rotating proton pumping ATPases. (2010)
  • Author(s): M.Nakanishi-Matsui, M.Sekiya, R.K.Nakamoto, M.Futai
  • Journal Title: Biochim.Biophys.Acta Volume: 1797 Pages: 1343-1352
  • Peer Reviewed
• [Journal Article] Roles of the β subunit hinge domain in ATP synthase F_1 sector : Hydrophobic network formed by introduced β Phe174 inhibits subunit rotation. (2010)
  • Author(s): M.Nakanishi-Matsui, S.Kashiwagi, M.Kojima, T.Nonaka, M.Futai
  • Journal Title: Biochem.Biophys.Res.Commun. Volume: 395 Pages: 173-177
  • Peer Reviewed
• [Journal Article] Single molecule behavior of inhibited and active States of Escherichia coli ATP synthase F_1 rotation. (2010)
  • Author(s): M.Sekiya, H.Hosokawa, M.Nakanishi-Matsui, M.K.Al-Shawi, R.K.Nakamoto, M.Futai
  • Journal Title: J.Biol.Chem. Volume: 285 Pages: 42058-42067
  • Peer Reviewed
• [Journal Article] Temperature dependence of single molecule rotation of the Escherichia coli ATP synthase F_1 sector reveals the importance of α・β subunit interactions in the catalytic dwell. (2009)
  • Author(s): M. Sekiya, R.K. Nakamoto, M.K. Al-Shawi, M. Nakanishi-Matsui, M. Futai
  • Journal Title: J.Biol.Chem. 284 Pages: 22401-22410
• [Journal Article] Temperature dependence of single molecule rotation of the Escherichiacoli ATP synthase F_1 sector reveals the importance of γ-β subunit interactions in the catalytic dwell. (2009)
  • Author(s): Mizuki Sekiya, Masamitsu Futai, et al
  • Journal Title: J.Biol. Chem. 284 Pages: 22401-22410
  • Peer Reviewed
• [Journal Article] Effects of mutations in β subunit hinge region on ATP synthase F_1 sector rotation. (2008)
  • Author(s): S. Kashiwagi, A. Iwamoto-Kihara, M. Nakanishi-Matsui, M. Futai
  • Journal Title: Biochem.Biophys.Res.Commun. 365 Pages: 227-231
• [Journal Article] The V-type H+-ATPase in vesicular trafficking : targeting, regulation and function. (2008)
  • Author(s): V. Marshansky, M. Futai
  • Journal Title: Curr.Opin.Cell Biol. 20 Pages: 415-426
• [Journal Article] Stochastic rotational catalysis of proton pumping F-ATPase. (2008)
  • Author(s): M. Nakanishi-Matsui, M. Futai
  • Journal Title: Phil.Trans.Royal Soc. B363 Pages: 2135-2142
• [Journal Article] Unique assembly of a hybrid yeast vacuolar H^+-ATPase containing mouse testis specific E1 isoform. (2008)
  • Author(s): K. Hayashi, G.H. Sun-Wada, Y. Wada, M. Nakanishi-Matsui, M. Futai
  • Journal Title: Biochim.Biophys.Acta 1777(10) Pages: 1370-1377
• [Journal Article] Effects of mutations in β subunit hinge region on ATP synthase F_1 sector rotation. (2008)
  • Author(s): S. Kashiwagi, et al.
  • Journal Title: Biochem. Biophys. Res. Commun. 365 Pages: 227-231
  • Peer Reviewed
• [Journal Article] The V-type H^+-ATPase in vesicular trafficking : targeting, regulation and function (2008)
  • Author(s): V. Marshansky, et. al.
  • Journal Title: Current Opinion in Cell Biology 20 Pages: 1-12
  • Peer Reviewed
• [Journal Article] Stochastic rotational catalysis of proton pumping F ATPase (2008)
  • Author(s): M. Nakanishi-Matsui, et al.
  • Journal Title: Phil. Trans. Royal Soc. B 363 Pages: 2135-2142
  • Peer Reviewed
• [Journal Article] Defective assembly of a hybrid vacuolar H(+)-ATPase containing the mouse testis-specific E1 isoform and yeast subunits (2008)
  • Author(s): K. Hayashi, et al.
  • Journal Title: Biochimica et Biophysica Acta 1777 Pages: 1370-1377
  • Peer Reviewed
• [Journal Article] The mechanism of rotating proton pumping ATPases.
  • Author(s): Mayumi Nakanishi-Matsui, Masamitsu Futai, et al.
  • Journal Title: Biochim.Biophys.Acta (In press)
  • Peer Reviewed
• [Journal Article] Roles of the β subunit hinge domain in ATP synthase F_1 sector : hydrophobic network formed by introduced β Phe174 inhibits subunit rotation.
  • Author(s): Mayumi Nakanishi-Matsui, Masamitsu Futai, et al.
  • Journal Title: Biochem. Biophys.Res.Commun. (In press)
  • Peer Reviewed
• [Presentation] 破骨細胞に発現するV-ATPaseのイソフォーム構成とATPase活性 (2011)
  • Author(s): 後藤(松元)奈緒美
  • Organizer: 日本薬学会第131年会
  • Place of Presentation: 静岡
  • Year and Date: 2011-03-30
• [Presentation] 大腸菌ATP合成酵素におけるεサブユニットの役割 (2011)
  • Author(s): 関谷瑞樹
  • Organizer: 日本薬学会第131年会
  • Place of Presentation: 静岡
  • Year and Date: 2011-03-30
• [Presentation] 破骨細胞に発現するV-ATPaseのイソフォーム構成とATPase活性 (2011)
  • Author(s): 松元奈緒美, 中西(松井)真弓, 二井將光
  • Organizer: 日本薬学会第131年会
  • Place of Presentation: 静岡((東北関東大震災のため中止) 当初の予定)
  • Year and Date: 2011-03-30
• [Presentation] 大腸菌ATP合成酵素におけるεサブユニットの役割 (2011)
  • Author(s): 関谷瑞樹, 中西(松井)真弓, 西山枝里, R.K.Nakamoto, M.K.Al-Shawi, 二井將光
  • Organizer: 日本薬学会第131年会
  • Place of Presentation: 静岡((東北関東大震災のため中止) 当初の予定)
  • Year and Date: 2011-03-30
• [Presentation] A functional analysis of vacuolar H+ ATPase subunit C in yeast (2010)
  • Author(s): 岡本晴子
  • Organizer: BMB2010
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-09
• [Presentation] Effect of ε subunit on F_1 rotation in E. coli ATP synthase (2010)
  • Author(s): 関谷瑞樹
  • Organizer: BMB2010
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-09
• [Presentation] Single molecule detection of an intrinsic inhibited state of the E. coli F_1 ATPase (2010)
  • Author(s): 関谷瑞樹
  • Organizer: BMB2010
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-09
• [Presentation] Single molecule detection of an intrinsic inhibited state of the E.coli F_1 ATPase (2010)
  • Author(s): 關谷瑞樹, 細川浩之, Robert K.Nakamoto, Marwan K.Al-Shawi, 中西(松井)真弓, 二井將光
  • Organizer: BMB2010
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-09
• [Presentation] Effect of ε subunit on F_1 rotation in E.coli ATP synthase. (2010)
  • Author(s): 關谷瑞樹, 西山枝里, R.K.Nakamoto, 中西(松井)真弓, 二井將光
  • Organizer: BMB2010
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-09
• [Presentation] Afunctional analysis of vacuolar H^+ ATPase subunit C in yeast. (2010)
  • Author(s): 岡本晴子, 二井將光
  • Organizer: BMB2010
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-09
• [Presentation] LPS(リポポリサッカライド)が誘導するマクロファージの細胞融合 (2010)
  • Author(s): 中西真弓
  • Organizer: BMB2010
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-07
• [Presentation] The intrinsic inhibition mechanism of ATP hydrolysis in ATP synthase : Single molecule observation. (2010)
  • Author(s): 関谷瑞樹
  • Organizer: FAOBMB Symposium (OzBio2010)
  • Place of Presentation: Melbourne
  • Year and Date: 2010-09-27
• [Presentation] The intrinsic inhibition mechanism of ATP hydrolysis in ATP synthase : Single molecule observation. (2010)
  • Author(s): M.Sekiya, H.Hosokawa, M.Nakanishi-Matsui, M.Futai
  • Organizer: FAOBMB Symposium(OzBio2010)
  • Place of Presentation: Melbourne, Australia
  • Year and Date: 2010-09-27
• [Presentation] Different localization of specific V-ATPase between osteoclast and LPS-induced multinuclear cells. (2010)
  • Author(s): M.Nakanishi-Matsui, S., Yano, N.Matsumoto, M.Futai
  • Organizer: FASEB Summer research conferences
  • Place of Presentation: Colorado, USA
  • Year and Date: 2010-06-10
• [Presentation] Thermodynamics of single molecule rotation of Escherichia coli F-ATPase. (2010)
  • Author(s): M.Sekiya, H.Hosokawa, R.K.Nakamoto, M.K.Al-Shawi, M.Nakanishi-Matsui, M.Futai
  • Organizer: FASEB Summer research conferences
  • Place of Presentation: Colorado, USA
  • Year and Date: 2010-06-09
• [Presentation] プロトンの生理学的役割について (2010)
  • Author(s): 二井將光
  • Organizer: 第87回日本生理学会
  • Place of Presentation: 岩手
  • Year and Date: 2010-05-21
• [Presentation] プロトンの生理学的役割について (2010)
  • Author(s): 二井將光
  • Organizer: 第87回日本生理学会
  • Place of Presentation: 盛岡, 岩手(記念講演)
  • Year and Date: 2010-05-21
• [Presentation] What we have learned from single molecules : from the results of F-ATPase (F_oF_1). (2010)
  • Author(s): 二井將光
  • Organizer: Symposium on The Future of Biochemistry and Molecular Biology in the 21 Century
  • Place of Presentation: Seoul National University, Seoul
  • Year and Date: 2010-05-08
• [Presentation] What we have learned from single molecules : from the results of F-ATPase(F_0F_1) (2010)
  • Author(s): 二井將光
  • Organizer: Symposium on The Future of Biochemistry and Molecular Biology in the 21 Century
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2010-05-08
• [Presentation] 大腸菌ATP合成酵素によるATP加水分解反応の阻害機構 (2010)
  • Author(s): 関谷瑞樹
  • Organizer: 日本薬学会第130年会
  • Place of Presentation: 就実大学(岡山)
  • Year and Date: 2010-03-29
• [Presentation] 大腸菌ATP合成酵素によるATP加水解反応の阻害機構 (2010)
  • Author(s): 關谷瑞樹, 中西(松井)真弓, 二井將光, 他
  • Organizer: 日本薬学会第130年会
  • Place of Presentation: 就実大学(岡山)
  • Year and Date: 2010-03-29
• [Presentation] 破骨細胞におけるV-ATPase a3サブユニット局在オルガネラの解析 (2009)
  • Author(s): 後藤(松元奈緒美)
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2009-10-23
• [Presentation] ATP合成酵素の回転における熱力学とγ-βサブユニット間の相互作用の重要性 (2009)
  • Author(s): 関谷瑞樹
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2009-10-22
• [Presentation] Pauses of subunit rotation of Escherichia coli ATP synthase F_1 Sector. (2009)
  • Author(s): 中西真弓
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2009-10-22
• [Presentation] Thermodynamics of single molecule rotation of proton pumping ATPase (2009)
  • Author(s): 関谷瑞樹
  • Organizer: 21st IUBMB and the 12th FAOBMB International Congress of Biochemistry and Molecular Biology
  • Place of Presentation: 上海国際会議センター
  • Year and Date: 2009-08-06
• [Presentation] Characterization of organelles with V-ATPase a3 subunit in osteoclasts (2009)
  • Author(s): 後藤(松元)奈緒美
  • Organizer: JBS Biofrontier Symposium on Biochemistry of pH Homeostasis and Proton Circuit
  • Place of Presentation: 岩手医科大学矢巾キャンパス
  • Year and Date: 2009-07-24
• [Presentation] Effect of magnetic fields on blue-light receptor, cryptochrome -dependent responses in Arabidopsis thaliana (2009)
  • Author(s): 岡本晴子
  • Organizer: JBS Biofrontier Symposium on Biochemistry of pH Homeostasis and Proton Circuit
  • Place of Presentation: 岩手医科大学矢巾キャンパス
  • Year and Date: 2009-07-24
• [Presentation] Thermodynamics of single molecule rotation of Escherichia coli ATP synthase. (2009)
  • Author(s): 関谷瑞樹
  • Organizer: JBS Biofrontier Symposium on Biochemistry of pH Homeostasis and Proton Circuit
  • Place of Presentation: 岩手医科大学矢巾キャンパス
  • Year and Date: 2009-07-24
• [Presentation] Subunit rotation of E. coli F1-ATPase : Effects of mutations in the β subunit hinge domain. (2009)
  • Author(s): 中西真弓
  • Organizer: JBS Biofrontier Symposium on Biochemistry of pH Homeostasis and Proton Circuit
  • Place of Presentation: 岩手医科大学矢巾キャンパス
  • Year and Date: 2009-07-24
• [Presentation] Subunit rotation of E. coli F_1-ATPase : Effects of mutations in the β subunit hing ε edomain. (2009)
  • Author(s): Mayumi Nakanishi-Matsui, Masamitsu Futai, et al.
  • Organizer: JBS Biofrontier Symposium on Biochemistry of pH Homeostasis and Proton Circuit
  • Place of Presentation: 岩手医科大学矢巾キャンパス
  • Year and Date: 2009-07-24
• [Presentation] Thermodynamics of Escherichia coli ATP synthase F_1 sector rotation and effects of γ subunit Met23 replacement. (2009)
  • Author(s): 関谷瑞樹
  • Organizer: Iwate International Symposium on Biochemical Approaches to Drug Discovery
  • Place of Presentation: 岩手医科大学矢巾キャンパス
  • Year and Date: 2009-05-15
• [Presentation] Mechanism of proton transport ATPases : rotation of ATP synthase F_1 sector. (2009)
  • Author(s): 中西真弓
  • Organizer: Iwate International Symposium on Biochemical Approaches to Drug Discovery
  • Place of Presentation: 岩手医科大学矢巾キャンパス
  • Year and Date: 2009-05-15
• [Presentation] ATP合成酵素の回転における熱力学とβ-γサブユニット間の相互作用による影響 (2009)
  • Author(s): 関谷瑞樹
  • Organizer: 生化学会東北支部会
  • Place of Presentation: 東北大学片平さくらホール
  • Year and Date: 2009-05-09
• [Presentation] ATP合成酵素の回転におけるβ-γサブユニット間相互作用の影響 (2009)
  • Author(s): 関谷瑞樹
  • Organizer: 日本薬学会第129年会
  • Place of Presentation: 京都
  • Year and Date: 2009-03-26
• [Presentation] ATP合成酵素の回転におけるβ-αサブユニット間相互作用の影響(口頭) (2009)
  • Author(s): 関谷瑞樹
  • Organizer: 日本薬学会 第129年会
  • Place of Presentation: 京都
  • Year and Date: 2009-03-26
• [Presentation] The effects of γ subunit Met23 replacement on ATP synthase F_1 sector rotation. (2008)
  • Author(s): 関谷瑞樹
  • Organizer: BMB2008
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-10
• [Presentation] Rotation of ATP synthase F_1 sector with mutations in β subunit hinge domain. (口頭及びポスター) (2008)
  • Author(s): 柏木幸子
  • Organizer: BMB2008(第31回日本分子生物学会年会・第81回日本生化学会大会合同大会)
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-10
• [Presentation] The effects of γ subunit Met23 replacement on ATP synthase F_1 sector rotation. (口頭) (2008)
  • Author(s): 関谷瑞樹
  • Organizer: BMB2008(第31回日本分子生物学会年会・第81回日本生化学会大会合同大会)
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-10
• [Presentation] ATP合成酵素のβ-γサブユニット間相互作用による回転への影響 (2008)
  • Author(s): 関谷瑞樹
  • Organizer: 第47回日本薬学会東北支部大会
  • Place of Presentation: 岩手
  • Year and Date: 2008-10-26
• [Presentation] ATP合成酵素の残基間相互作用と回転触媒機構(口頭) (2008)
  • Author(s): 柏木幸子
  • Organizer: 第47回日本薬学会 東北支部大会
  • Place of Presentation: 岩手
  • Year and Date: 2008-10-26
• [Presentation] ATP合成酵素のβ-γサブユニット間相互作用による回転への影響(口頭) (2008)
  • Author(s): 関谷瑞樹
  • Organizer: 第47回日本薬学会 東北支部大会
  • Place of Presentation: 岩手
  • Year and Date: 2008-10-26
• [Presentation] Effects of mutations in the β subunit hinge domain on ATP synthase F_1 sector rotation. (2008)
  • Author(s): 中西真弓
  • Organizer: Mini -Symposium on Proton Pumping ATPases-Iwate-Oxford Collaboration
  • Place of Presentation: 岩手医科大学
  • Year and Date: 2008-09-22
• [Presentation] The effect of γ subunit Met23 replacement on ATP synthase F1 sector rotation. (2008)
  • Author(s): 関谷瑞樹
  • Organizer: Mini-Symposium on Proton Pumping ATPases-Iwate-Oxford Collaboration
  • Place of Presentation: 岩手医科大学
  • Year and Date: 2008-09-22
• [Presentation] Rotation of ATP synthase F_1 sector with mutations in β subunit hinge domain. (2008)
  • Author(s): 中西真弓
  • Organizer: Bio Nano Technology
  • Place of Presentation: 神戸
  • Year and Date: 2008-09-17
• [Presentation] Rotation of ATP synthase F_1 sector with mutations in β subunit hinge domain. (口頭) (2008)
  • Author(s): 中西真弓
  • Organizer: Bio Nano Technology
  • Place of Presentation: 神戸
  • Year and Date: 2008-09-17
• [Presentation] Effects of mutations in the beta subunit hinge domain on ATP synthase F_1 sector rotation. (2008)
  • Author(s): M. Futai
  • Organizer: 33^<rd> FEBS Congress and 11^<th> IUBMB Conference : Biochemistry of Cell Regulation
  • Place of Presentation: アテネ
  • Year and Date: 2008-07-01
• [Presentation] Effects of mutations in the beta subunit hinge domain on ATP synthase F_1 sector rotation (口頭) (2008)
  • Author(s): 二井将光
  • Organizer: 33^<rd> FEBS Congress and 11^th IUBMB Conference : Biochemistry of Cell Regulation
  • Place of Presentation: アテネ
  • Year and Date: 2008-07-01
• [Presentation] Stochastic rotational catalysis of proton pumping F-ATPase. (2008)
  • Author(s): M. Futai
  • Organizer: Royal Society and JSPS Joint Symposium
  • Place of Presentation: ロンドン 招待講演
  • Year and Date: 2008-06-15
• [Presentation] Single-Molecule Imaging (口頭) (2008)
  • Author(s): 二井将光
  • Organizer: Royal Society and JSPS Joint Symposium
  • Place of Presentation: ロンドン
  • Year and Date: 2008-06-15
• [Presentation] Different localization of specific V-ATPase between osteoclast and LPS-induced multinuclear cells.
  • Author(s): 中西真弓
  • Organizer: FASEB Summer research conferences
  • Place of Presentation: Colorado, USA
• [Presentation] Thermodynamics of single molecule rotation of Escherichia coli F-ATPase.
  • Author(s): 関谷瑞樹
  • Organizer: FASEB Summer research conferences
  • Place of Presentation: Colorado, USA
• [Book] 科学 (2008)
  • Author(s): 二井将光
  • Total Pages: 115
  • Publisher: 岩波書店
• [Remarks] ホームページ等