Elucidation of the structural basis of the excitation and contraction coupling usiing phase-contrast electron cryotomography

• Project/Area Number: 23570150
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: National Institute for Physiological Sciences
• Principal Investigator: MURATA Kazuyoshi 生理学研究所, 脳機能計測・支援センター, 准教授 (20311201)
• Co-Investigator(Kenkyū-buntansha): KUNIYASU Akihiko 崇城大学, 薬学部, 教授 (90241348)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2012: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2011: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 高分解能電子顕微鏡解析 / 膜タンパク質 / 興奮収縮連関 / クライオ電子顕微鏡 / 電子線トモグラフィー

Research Abstract

We extracted enzymatically-active triad membranes from rabbit's skeletal muscle and examined them using electron cryomicroscopy. A lot of proteoliposomes were successfully observed in the sample. Some of them were identified as sarcoplasmic reticulum (SR). We further labeled the dihydropyridine receptor (DHPR) in T-tubules using the anti-beta subunit antibody conjugated fluorescent dyes and gold colloids to detect the triad junction (TJ). Although we can find gold-labels near SRs, The region of TJ has not been found out yet. We will also reconsider the condition of membrane extraction to preserve TJ abundantly, and examine it again in the future.

Research Products

• [Journal Article] クライオ位相差電顕トモグラフィーによる興奮収縮連関の構造的基盤の解明 (2013)
  • Author(s): 村田和義、國安明彦
  • Journal Title: 生理学研究所年報 Volume: 34 Pages: 80-81
• [Journal Article] 骨格筋トライアドジャンクションの三次元構造解析 (2013)
  • Author(s): 村田和義、國安明彦
  • Journal Title: 生理学研究所年報 Volume: 33 Pages: 195-196
• [Journal Article] An archaeal homolog of proteasome assembly factor functions as a proteasome activator (2013)
  • Author(s): Kumoi K, Satoh T, Murata K, Hiromoto T, Mizushima T, Kamiya Y, Noda M, Uchiyama S, Yagi H & Kato K.
  • Journal Title: PloS one Volume: 8
  • Peer Reviewed
• [Journal Article] Three-dimensional evaluation of the spinal local neural network revealed by the high-voltage electron microscopy: a double immunohistochemical study (2012)
  • Author(s): Oti T, Satoh K, Saito K, Murata K, Kawata M, Sakamoto T & Sakamoto H.
  • Journal Title: Histochemistry and cell biology Volume: 138 Pages: 693-697
  • Peer Reviewed
• [Journal Article] Structural basis for broad detection of genogroup II noroviruses by a monoclonal antibody that binds to a site occluded in the viral particle (2012)
  • Author(s): Hansman G, Taylor D, McLellan J, Smith T, Georgiev I, Tame J, Park SY, Yamazaki M, Gondaira F, Miki M, Katayama K, Murata K, Kwong P
  • Journal Title: Journal of Virology Volume: 86 Pages: 3635-3646
  • Peer Reviewed
• [Journal Article] 骨格筋トライアドジャンクションの三次元構造解析 (2011)
  • Author(s): 村田和義、國安明彦
  • Journal Title: 生理学研究所年報 Volume: 32 Pages: 210-210
• [Journal Article] Seeing the portal in herpes simplex virus type 1 B capsids (2011)
  • Author(s): Rochat RH, Liu X, Murata K, Nagayama K, Rixon FJ, Chiu W
  • Journal Title: Journal of Virology Volume: 85 Pages: 1871-1874
  • Peer Reviewed
• [Presentation] クライオ超高圧電子顕微鏡トモグラフィーによるシアノバクテリアDNA超構造の観察
  • Author(s): 村田和義、萩原小百合、関由起子、金子康子
  • Organizer: 日本顕微鏡学会
  • Place of Presentation: 茨城
• [Remarks] 研究概要 ハイライト：
  • URL: http://www.nips.ac.jp/struct/overviews-j.html