Structural studied of Complex I and V-ATPase by Electron microscopy

• Project/Area Number: 21370042
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Kyoto Sangyo University (2010-2011); Tokyo Institute of Technology (2009)
• Principal Investigator: YOKOYAMA Ken 京都産業大学, 総合生命科学部, 教授 (70271377)
• Co-Investigator(Kenkyū-buntansha): MITSUOKA Kaoru 産業技術総合研究所, バイオメディシナル情報研究センター, 研究チーム長 (60301230)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥18,850,000 (Direct Cost: ¥14,500,000、Indirect Cost: ¥4,350,000); Fiscal Year 2011: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000); Fiscal Year 2010: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2009: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
• Keywords: ATP / 呼吸 / 構造 / 活性酸素 / 電子顕微鏡

Research Abstract

We have prepared a 2-D crystal of rotor ring from VoV1. We are reconstituting 3D model of the rotor ring from images by cryo electrol microscopy. For Complex I, his-tagged purification system was constructed using homologus recombination method. Single particle analysis revealed the obtained Complex I is intact.

Research Products

• [Journal Article] Expression of ATP sensor protein in Caenorhabditis elegans (2012)
  • Author(s): J. Kishikawa, M. Fujikawa, H. Imamura, K. Yasuda, H. Noji, N. Ishii, S. Mitani, and K. Yokoyama
  • Journal Title: Microsc. Res. Tech Volume: 75(1) Issue: 1 Pages: 15-9
  • DOI: 10.1002/jemt.21103
  • Peer Reviewed
• [Journal Article] Expression of ATP sensor protein in Caenorhabditis elegans、Microsc (2012)
  • Author(s): J.Kishikawa, M.Fujikawa, H.Imamura, K.Yasuda, H.Noji, N.Ishii, S.Mitani, K.Yokoyama
  • Journal Title: Res.Tech Volume: (未定)(in press)
  • Peer Reviewed
• [Journal Article] Resolving stepping rotation in Thermus thermophilis H+-ATPase/synthase with an essentially drag-free probe (2011)
  • Author(s): Furuike, S., et al.
  • Journal Title: Nat. Commun. Volume: 2 Issue: 1 Pages: 223-239
  • DOI: 10.1038/ncomms1215
  • Peer Reviewed
• [Journal Article] Resolving stepping rotation in Thermus thermophilus H^+-ATPase/synthase with an essentially drag free probe (2011)
  • Author(s): Furuike, S, Nakano, M., Adachi, K., Noji, H., Kinosita Jr., K., Yokoyama, K
  • Journal Title: Nature Communications Volume: 2 Pages: 233-239
  • Peer Reviewed
• [Journal Article] Mechanism of inhibition of Tributyltin chloride of the V type Molecular Motor (2009)
  • Author(s): Takeda M, Ikeda C, Shimabukuro K, Yoshida M, Yokoyama K
  • Journal Title: Biophysical J Volume: Vol 96 Issue: 3 Pages: 1210-1217
  • DOI: 10.1016/j.bpj.2008.10.031
  • Peer Reviewed
• [Journal Article] K. Cryo-EM structure of the native GroEL-GroES complex from thermos thermophilus encapsulating substrate inside the cavity (2009)
  • Author(s): K, Taguchi、H, Mitsuok
  • Journal Title: Structure Volume: Vol18 Pages: 287-293
  • Peer Reviewed
• [Journal Article] Cryo-EM structure of the native GroEL-GroES complex from thermos thermophilus encapsulating substrate inside the cavity (2009)
  • Author(s): Kanno R, Koike-Takeshita A, Yokoyama K, Taguchi H, Mitsuoka K
  • Journal Title: Structure Volume: Vol18 Issue: 2 Pages: 287-293
  • DOI: 10.1016/j.str.2008.12.012
  • Peer Reviewed
• [Journal Article] Crystal structure of A3B3 compiex of V-ATP from Thermus thermophilus (2009)
  • Author(s): Maher MJ, Akimoto S, Iwata M, Nagata K, Hori Y, Yoshida M, Yokoyama S, Iwata S, Yokoyama K
  • Journal Title: EMBO J Volume: Vol28 Issue: 23 Pages: 3771-3779
  • DOI: 10.1038/emboj.2009.310
  • Peer Reviewed
• [Journal Article] Crystal structure of A3B3 complex of V-ATPase from Thermus thermophilus. (2009)
  • Author(s): Maher MJ, Akimoto S, Iwata M, Nagata K, Hori Y, Yoshida M, Yokoyama S, Iwata S, Yokoyama K.
  • Journal Title: EMBO J Vol 28 Pages: 3771-3779
  • Peer Reviewed
• [Journal Article] Cryo-EM structure of the native GroEL-GroES complex from thermus thermophilus encapsulating substrate inside the cavity. (2009)
  • Author(s): Kanno R, Koike-Takeshita A, Yokoyama K, Taguchi H, Mitsuoka K.
  • Journal Title: Structure Vol 28 Pages: 287-293
  • Peer Reviewed
• [Journal Article] Mechanism of inhibition of Tributyltin chloride of the V type Molecular Motor. (2009)
  • Author(s): Takeda M, Ikeda C, Shimabukuro K, Yoshida M, Yokoyama K.
  • Journal Title: Biophysical J Vol 96 Pages: 1210-1217
  • Peer Reviewed
• [Presentation] 回転分子モーターの軸サブユニットの分子進化 (2011)
  • Author(s): 岸川淳一, 伊吹達也, 南野徹, 難波啓一, 中西温子, 宮田知子, 今田勝巳, 横山謙
  • Organizer: 第37回日本生体エネルギー研究会討論会
  • Place of Presentation: 京都市
  • Year and Date: 2011-12-20
• [Presentation] 回転分子モーターの軸サブユニットの分子進化 (2011)
  • Author(s): 岸川淳一, 伊吹達也, 南野徹, 難波啓一, 中西温子, 宮田知子, 今田勝巳, 横山謙
  • Organizer: 37回日本生体エネルギー研究会討論会
  • Place of Presentation: 京都市
  • Year and Date: 2011-12-20
• [Presentation] ATP concentration change in Ca enorhabditis elegans (2011)
  • Author(s): J. Kishikawa, M. Fujikawa, H. Imamura, K. Yasuda, H. Noji, S. Mitani, and K. Yokoyama
  • Organizer: The 34th annual me eting of the molecular biology society of Japan
  • Place of Presentation: Yokohama
  • Year and Date: 2011-12-14
• [Presentation] ATP concentration change in Caenorhabditis elegans (2011)
  • Author(s): J.Kishikawa, M.Fujikawa, H.Imamura, K.Yasuda, H.Noji, S.Mitani, K.Yokoyama
  • Organizer: The 34th annual meeting of the molecular biology society of Japan
  • Place of Presentation: 横浜市
  • Year and Date: 2011-12-14
• [Presentation] 無負荷回転プローブによる好熱菌VoV_1/V_1のステップ解析 (2010)
  • Author(s): 古池晶, 木下一彦, 横山謙
  • Organizer: 36回日本生体エネルギー研究会
  • Place of Presentation: 大阪
  • Year and Date: 2010-11-19
• [Presentation] 無負荷回転プローブによる好熱菌VoV_1/V_1のステップ解析 (2010)
  • Author(s): 古池晶, 木下一彦, 横山謙
  • Organizer: 36回日本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] Resolving s tepping rotation of V-ATPase with an es sentially drag-free probe (2010)
  • Author(s): Furuike S, Nakano M, Adachi K, Noji H, Kinoshita K Jr, Yokoyama K
  • Organizer: 16th European Bioenegetics conference
  • Place of Presentation: ポーランド
  • Year and Date: 2010-07-18
• [Presentation] Resolving stepping rotation of V-ATPase with an essentially drag-free probe (2010)
  • Author(s): Furuike S., Nakano M., Adachi K., Noji H., Kinoshita K.Jr., Yokoyama K.
  • Organizer: 16th European Bioenegetics conference
  • Place of Presentation: ポーランド(ワルシャワ)
  • Year and Date: 2010-07-18
• [Remarks]