Genetic approach on subunit architecture of sodium-translocating V-ATPase complex

• Project/Area Number: 21570144
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: Ehime University
• Principal Investigator: KAKINUMA Yoshimi 愛媛大学, 農学部, 教授 (80134394)
• Co-Investigator(Renkei-kenkyūsha): MURATA Takeshi 千葉大学, 理学部, 特任准教授 (80415322)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2010: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2009: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: ナトリウム / V. ATPアーゼ / 腸球菌 / イオン / サブユニット / V-APTアーゼ / V-ATPアーゼ

Research Abstract

Enterococcus hirae V-ATPase is an ion-transporting rotary enzyme that consists of nine subunits. In order to clarify the molecular mechanism of this enzyme, it is essential to clarify the overall architecture and the interaction among subunits. In this study, reconstruction of V-ATPase complex was succeeded from each subunit. Based on crystal structure of the subunit complexes and directed mutagenesis study, the features of individual V-ATPase subunits was revealed.

Research Products

• [Journal Article] Crystal structure of the central axis DF complex of the prokaryotic V-ATPase (2011)
  • Author(s): Saijo S, Arai S, Hossain KM, Yamato I, Suzuki K, Kakinuma Y, Ishizuka-Katsura Y, Ohsawa N, Terada T, Shirouzu M, Yokoyama S, Iwata S, Murata T
  • Journal Title: Proc Natl Acad Sci USA Volume: 108(50) Pages: 19955-19960
  • Peer Reviewed
• [Journal Article] Structure of the rotor ring modified with N, N'-dicyclohexylcarbodiimide of the Na+-transporting vacuolar ATPase (2011)
  • Author(s): Mizutani K, Yamamoto M, Suzuki K, Yamato I, Kakinuma Y, Shirouzu M, Walker JE, Yokoyama S, Iwata S, Murata T
  • Journal Title: Proc Natl Acad Sci USA Volume: 108(33) Pages: 13474-13479
  • Peer Reviewed
• [Journal Article] Expression and purification of the central stalk subunits of Na+-translocating V-type ATPase from Enterococcus hirae (2011)
  • Author(s): K. M. Mozaffor Hossain, Satoshi Arai, Shinya Saijo, Yoshimi Kakinuma, Takeshi Murata, and Ichiro Yamato
  • Journal Title: African Journal of Biotechnology Volume: Vol.10(13) Pages: 2406-2413
  • Peer Reviewed
• [Journal Article] Significance of the glutamate-139 residue of the V-type Na+-ATPase NtpK subunit in catalytic turnover linked with salt tolerance of Enterococcus hirae (2011)
  • Author(s): Kawano-Kawada M, Takahashi H, Igarashi K, Murata T, Yamato I, Homma M, Kakinuma Y
  • Journal Title: J Bacteriol Volume: 193(14) Pages: 3657-3661
  • Peer Reviewed
• [Journal Article] Crystal structure of the central axis DF complex of the prokaryotic V-ATPase (2011)
  • Author(s): Saijo, S.
  • Journal Title: Pioc Natl Acad Sci USA Volume: 108 Issue: 50 Pages: 19955-19960
  • DOI: 10.1073/pnas.1108810108
  • Peer Reviewed
• [Journal Article] Structure of the rotor ring modified with N,N'-dicyclohexylcarbodiimide of the Na+-transporting vacuolar ATPase (2011)
  • Author(s): Mizutani, K.
  • Journal Title: Proc Natl Acad Sci USA Volume: 108 Issue: 33 Pages: 13474-13479
  • DOI: 10.1073/pnas.1103287108
  • Peer Reviewed
• [Journal Article] Significance of the glutamate-139 residue of the V-type Na+-ATPase NtpK subunit in catalytic turnover linked with salt tolerance of Enterococcus hirae (2011)
  • Author(s): Kawano-Kawada, M.
  • Journal Title: Journal of Bacteriology Volume: 193 Issue: 14 Pages: 3657-3661
  • DOI: 10.1128/jb.01537-10
  • Peer Reviewed
• [Journal Article] Reconstitution in vitro of the catalytic portion(NtpA3-B3-D-G complex) of Enterococcus hirae V-type Na+-ATPase (2009)
  • Author(s): Arai S, Yamato I, Shiokawa A, Saijo S, Kakinuma Y, Ishizuka-Katsura Y, Toyama M, Terada T, Shirouzu M, Yokoyama S, Iwata S, Murata T
  • Journal Title: Biochem Biophys Res Commun Volume: 390(3) Pages: 698-702
  • Peer Reviewed
• [Journal Article] Tributyltin sensitivity of vacuolar-type Na(+)-transporting ATPase from Enterococcus hirae (2009)
  • Author(s): Chardwiriyapreecha S, Inoue T, Sugimoto N, Sekito T, Yamato I, Murata T, Homma M, Kakinuma Y
  • Journal Title: J Toxicol Sci Volume: 34(5) Pages: 575-579
  • NAID: 110007358569
  • Peer Reviewed
• [Journal Article] Reconstitution in vitro of the catalytic portion(NtpA3-B3-D-G complex)of Enterococcus hirae V-type Na+-ATPase (2009)
  • Author(s): Arai, S.
  • Journal Title: Biochem Biophys Res Commun 390(3) Pages: 698-702
  • Peer Reviewed
• [Journal Article] Tributyltin sensitivity of vacuolar-type Na(+)-transporting ATPase from Enterococcus hirae (2009)
  • Author(s): Chardwiriyapreecha, S.
  • Journal Title: J Toxicol Sci. 34(5) Pages: 575-579
  • Peer Reviewed
• [Presentation] Mutagenesis of the residues forming ion-binding pocket of NtpK subunit of Enterococcus hirae V-ATPase (2011)
  • Author(s): Miyuki Kawano-Kawada, Tomoki Iwaki, Toshiaki Hosaka, Takeshi Murata, Ichiro Yamato, Michio Homma, Yoshimi Kakinuma
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2011-12-15
• [Presentation] Enterococcus hirae由来V_1-ATPaseの結晶構造解析 (2011)
  • Author(s): 西條慎也, 新井聡史, 山登一郎, 柿沼喜己, 石塚芳子, 寺田貴帆, 白水美香子, 横山茂之, 岩田想, 村田武士
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館
  • Year and Date: 2011-09-22
• [Presentation] 腸内連鎖球菌由来V-ATPaseのNtpA_3-B_3複合体の構造と非対称性形成機構 (2011)
  • Author(s): 新井聡史, 西條慎也, 山登一郎, 柿沼喜己, 石塚芳子, 寺田貴帆, 白水美香子, 横山茂之, 岩田想, 村田武士
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館
  • Year and Date: 2011-09-22
• [Presentation] Structure of the Na^+ unbound rotor ring modified with N, N'-Dicyclohexylcarbodiimide of the Na^+-transporting V-ATPase (2010)
  • Author(s): 水谷健二, 山本三沙岐, 山登一郎, 柿沼喜己, 本間道夫, 白水美香子, 横山茂之, 岩田想, 村田武士
  • Organizer: 第33回日本分子生物学会年会・第83回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-09
• [Presentation] V型ATPaseのサブユニット集合機構 (2010)
  • Author(s): 山本三沙岐, 水谷健二, 山登一郎, 柿沼喜己, 石塚芳子, 寺田貴帆, 白水美香子, 横山茂之, 岩田想, 村田武士
  • Organizer: 第33回日本分子生物学会年会・第83回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-09
• [Presentation] Subunit interactions and assembly of the V_1V_0-ATPase in vitro (2010)
  • Author(s): Yamamoto, M.
  • Organizer: 第33回日本分子生物学会年会・第83回日本生化学会大会
  • Place of Presentation: 神戸市
  • Year and Date: 2010-12-09
• [Presentation] Structure of the Na^+ unbound rotor ring modified with N, N'-Dicyclohexylcarbodiimide of the Na^+-transporting V-ATPase (2010)
  • Author(s): Mizutani, K.
  • Organizer: 第33回日本分子生物学会年会・第83回日本生化学会大会
  • Place of Presentation: 神戸市
  • Year and Date: 2010-12-09
• [Presentation] 腸内連鎖菌由来V_1-ATPaseのアセンブリーとNtpA_1-B_1中間複合体の構造解析 (2010)
  • Author(s): 新井聡史, 西條慎也, 山登一郎, 柿沼喜己, 石塚芳子, 寺田貴帆, 白水美香子, 横山茂之, 岩田想, 村田武士
  • Organizer: 第33回日本分子生物学会年会・第83回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-08
• [Presentation] Assembly mechanism of V_1-ATPase derived from Enterococcus hirae and crystallographic analysis of its NtpA_1-B_1 intermediate complex. (2010)
  • Author(s): Arai.S
  • Organizer: 第33回日本分子生物学会年会・第83回日本生化学会大会
  • Place of Presentation: 神戸市
  • Year and Date: 2010-12-08
• [Presentation] V1-ATPaseの発現精製とX線結晶構造解析 (2010)
  • Author(s): 村田武士、西條慎也、新井聡史、鈴木花野、水谷健二、石塚芳子、寺田貴帆、白水美香子、柿沼喜己、山登一郎、横山茂之、岩田想
  • Organizer: 第36回生体エネルギー研究会
  • Place of Presentation: 大阪大学銀杏会館
  • Year and Date: 2010-11-19
• [Presentation] 阻害剤DCCDが結合したV-ATPaseローターリングの構造 (2010)
  • Author(s): 水谷健二、山本三沙岐、山登一郎、柿沼喜己、白水美香子、John E. Walker、横山茂之、岩田想、村田武士
  • Organizer: 第36回生体エネルギー研究会
  • Place of Presentation: 大阪大学銀杏会館
  • Year and Date: 2010-11-18
• [Presentation] V型ATPaseのサブユニット集合機構 (2010)
  • Author(s): 山本三沙岐、水谷健二、山登一郎、柿沼喜己、石塚芳子、寺田貴帆、白水美香子、横山茂之、岩田想、村田武士
  • Organizer: 第36回生体エネルギー研究会
  • Place of Presentation: 大阪大学銀杏会館
  • Year and Date: 2010-11-18
• [Presentation] 腸内連鎖球菌V型ATPaseの膜内在性サブユニット(NtpI)の発現と精製 (2009)
  • Author(s): 山本三沙岐、水谷健二、柿沼喜己、山登一郎、岩田想、村田武士
  • Organizer: 第47回日本生物物理学会年会
  • Place of Presentation: アスティ徳島
  • Year and Date: 2009-11-01
• [Presentation] Expression and purification of the membrane subunit NtpI of Enterococcus hirae V-ATPase (2009)
  • Author(s): Yamamoto, M.
  • Organizer: 第47回日本生物物理学会年会
  • Place of Presentation: 徳島市
  • Year and Date: 2009-11-01