Elucidation of the intracellular protein folding mechanism?In vivo protein interaction analysis?

• Project/Area Number: 22570146
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: Kanagawa Institute of Technology
• Principal Investigator: KOIKE Ayumi 神奈川工科大学, 応用バイオ科学部, 教授 (20454176)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2012: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2011: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2010: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
• Keywords: タンパク質フォールディング / シャペロニン / GroEL / フォールディング / GroES / 相互作用 / タンパク質

Research Abstract

To elucidate the reaction mechanisms of GroEL, important in protein folding in the cell, I analyzed the GroEL / GroES interactions in cells using in vivo photo-crosslinking techniques. After cross-linking of the GroEL-GroES complexes formed in cells,they were analyzed by western blotting. As a result, “football” type reaction intermediate was detected. Furthermore, fluorescence correlation spectroscopy (FCS) using a GroES which fused fluorescent protein enabled to detect GroEL-GroES complexes in the cell.

Research Products

• [Journal Article] 新規ネガティブ染 色剤を用いた透過型電子顕微鏡による生物試 料の観察--分析電子顕微鏡システム利用研究成果 (2013)
  • Author(s): 依田ひろみ,小池あゆみ
  • Journal Title: 神奈川工科大学研究報告B理工学編 Volume: 37 Pages: 25-28
  • Peer Reviewed
• [Journal Article] 新規ネガティブ染色剤を用いた透過型電子顕微鏡による生物試料の観察 (2013)
  • Author(s): 依田ひろみ
  • Journal Title: 神奈川工科大学研究報告 Ｂ 理工学編 Volume: 37 Pages: 25-28
  • Peer Reviewed
• [Journal Article] Yeast prion protein New1 can break Sup35 amyloid fibrils into fragments in an ATP-dependent manner. (2010)
  • Author(s): Inoue Y, Kawai-Noma S, Koike-Takeshita A, Taguchi H, YoshidaM.
  • Journal Title: Genes Cells Volume: 16 Pages: 545-56
  • Peer Reviewed
• [Journal Article] Yeast prion protein New1 can break Sup35 amyloid fibrils into fragments in an ATP-dependent manner. (2010)
  • Author(s): Inoue Y, Kawai-Noma S, Koike-Takeshita A, Taguchi H, Yoshida M.
  • Journal Title: Genes Cells Volume: 16 Pages: 545-556
  • Peer Reviewed
• [Presentation] ナノサイズ図4GFP融合GroES発現系の空洞を持つ大腸菌シャペロニンGroELの水溶性キャッピング剤としての利用 (2012)
  • Author(s): 依田ひろみ,小池あゆみ
  • Organizer: 平成24年度神奈川県ものづくり技術交流会
  • Place of Presentation: 神奈川県産業 技術センター(海老名市)ポスター賞受賞
  • Year and Date: 2012-11-07
• [Presentation] シャペロニンGroEL変異体のナノ粒子キャッピング剤としての利用 (2012)
  • Author(s): 依田ひろみ,山本修,小池あゆみ
  • Organizer: 日本生物工学会
  • Place of Presentation: 神戸国際会議場(神戸市)
  • Year and Date: 2012-10-26
• [Presentation] GroEL mutant can encapsulates metal nano particles in each cavity (2012)
  • Author(s): Hiromi Yoda, Osamu Yamamoto, Ayumi Koike-Takeshita
  • Organizer: Pacific Rim Meeting
  • Place of Presentation: Hawaii、Honolulu
  • Year and Date: 2012-10-08
• [Presentation] 耐熱性GFPを用いた4型線毛関連タンパク質のThermus thermophilus内での可視化 (2011)
  • Author(s): 宮下沙也加
  • 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: 2011-12-15
• [Presentation] Visualization of the TypeIV pili proteins, PilD and PilT, tagged with superfolder green fluorescent protein in Thermus thermophilus (2011)
  • Author(s): Sayaka Miyashita
  • Organizer: 第1回モデル生物丸ごと一匹学会
  • Place of Presentation: Spring8(兵庫県)
  • Year and Date: 2011-08-20
• [Presentation] ASP-52 plays a critical role in ATP hydrolysis of GroEL (2011)
  • Author(s): Ayumi Koike-Takeshita
  • Organizer: Cold Spring Harbor Asia Conference"Protein Homeostasis in Health & Disease"
  • Place of Presentation: Suzhou Dushu Lake Conference Center(中国上海)
• [Presentation] In vivo evidence for the symmetric chaperonin GroEL-GroES complex revealed by site-directed in vivo photo-crosslinking. (2010)
  • Author(s): Ayumi Koike-Takeshita, Yukinori Yokota, Maki Okano , Megumi Masuda, Hideki Taguchi
  • Organizer: 日本分子生物学会
  • Place of Presentation: 神戸ポート アイランド(神戸市)
  • Year and Date: 2010-12-09
• [Presentation] In vivo evidence for the symmetric chaperonin GroEL-GroES complex revealed by site-directed in vivo photo-crosslinking. (2010)
  • Author(s): Ayumi Koike-Takeshita, Yukinori Yokota, Maki Okano, Megumi Masuda, Hideki Taguchi
  • Organizer: 日本分子生物学会
  • Place of Presentation: 神戸ポートアイランド(神戸市)
  • Year and Date: 2010-12-09
• [Presentation] 細胞内光クロスリンク技術を用いたシャペロニン複合体の解析 (2010)
  • Author(s): 小池あゆみ,横田幸範,岡野舞姫,増田恵,田口英樹
  • Organizer: 日本蛋白質科学会
  • Place of Presentation: 札幌コンベンションセンター(札幌市)
  • Year and Date: 2010-06-18
• [Presentation] ASP-52 plays a critical role in ATP hydrolysis of GroELCold Spring Harbor Asia Conference (2010)
  • Author(s): Ayumi Koike-Takeshita, Masasuke Yoshida, Hideki Taguchi
  • Organizer: Protein Homeostasis in Health & Disease
  • Place of Presentation: Suzhou Dushu Lake Conference Center(中国・上海)
• [Presentation] Analyses of interacting proteins with PilF and PilT from Thermus thermophilus.
  • Author(s): 安田卓人
  • Organizer: モデル生物丸ごと一匹学会
  • Place of Presentation: Spring-8（兵庫県）
• [Presentation] GroEL mutant can encapsulates metal nano particles in each cavity
  • Author(s): Hiromi Yoda
  • Organizer: Pacific Rim Meeting
  • Place of Presentation: Hawaii Convention Center and the Hilton Hawaiian Village (Honolulu, Hawaii)
• [Presentation] シャペロニンGroEL変異体のナノ粒子キャッピング剤としての利用
  • Author(s): 依田ひろみ
  • Organizer: 日本生物工学会
  • Place of Presentation: 神戸国際会議場（兵庫県）
• [Presentation] Detection of Amyoid Sup35NM Depending on Its Growth Using Field Effect Transistor
  • Author(s): Shofarul Wustoni
  • Organizer: 電気化学会
  • Place of Presentation: 東北大学（宮城県）
• [Presentation] シャペロニンGroEL/GroESの細胞内動態観察
  • Author(s): 小池あゆみ
  • Organizer: 日本蛋白質科学会
  • Place of Presentation: とりぎん文化会館（鳥取県）
• [Remarks]
  • URL: http://www.kanagawa-it.ac.jp/~l4013/syo/kyo%20gyoseki.pdf
• [Patent(Industrial Property Rights)] シャペロニン複合体及びその製造方法 (2012)
  • Inventor(s): 小池あゆみ、依田ひろみ、山本修
  • Industrial Property Rights Holder: 学校法人幾徳学園神奈川工科大学
  • Industrial Property Number: 2012-070329
  • Filing Date: 2012-03-26
• [Patent(Industrial Property Rights)] シャペロニン複合体及びその製造方法 (2012)
  • Inventor(s): 小池あゆみ
  • Industrial Property Rights Holder: 学校法人幾徳学園
  • Industrial Property Number: 2012-070329
  • Filing Date: 2012-03-26