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Development of methods to visualize small conformational change of proteins inside cells

Research Project

Project/Area Number 24657101
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeSingle-year Grants
Research Field Biophysics
Research InstitutionKyoto University

Principal Investigator

IMAMURA Hiromi  京都大学, 白眉センター, 准教授 (20422545)

Project Period (FY) 2012-04-01 – 2014-03-31
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2013: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2012: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywordsバイオイメージング / FRET / タンパク質動態 / バイオセンサー / 抗体 / アセチルCoA / 蛍光タンパク質 / イメージング
Research Abstract

Forster resonance energy transfer (FRET)-based biosensors that employ conformational changes of proteins are useful tools to obtain information inside living cells. However, it has been quite difficult to use proteins with small conformational changes for FRET biosensors. In this study, we used antibody to discriminate small conformational changes of proteins in order to overcome the above problem. We developed a FRET biosensor for an important metabolite, acetyl-CoA, by using an acetyl-CoA binding protein and an antibody against it.

Report

(3 results)
  • 2013 Annual Research Report   Final Research Report ( PDF )
  • 2012 Research-status Report
  • Research Products

    (29 results)

All 2014 2013 2012 Other

All Journal Article (11 results) (of which Peer Reviewed: 11 results,  Open Access: 3 results,  Acknowledgement Compliant: 1 results) Presentation (14 results) (of which Invited: 3 results) Book (1 results) Remarks (3 results)

  • [Journal Article] Toll-like receptor 9 protects non-immune cells from stress by modulating mitochondrial ATP synthesis through the inhibition of SERCA2.2014

    • Author(s)
      Shintani Y, Drexler HC, Kioka H, Terracciano CM, Coppen SR, Imamura H, Akao M, Nakai J, Wheeler AP, Higo S, Nakayama H, Takashima S, Yashiro K, Suzuki K.
    • Journal Title

      EMBO Reports

      Volume: 15 Issue: 4 Pages: 438-445

    • DOI

      10.1002/embr.201337945

    • Related Report
      2013 Final Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] ATP increases within the lumen of the endoplasmic reticulum upon intracellular Ca2+-release2014

    • Author(s)
      Vishnu, N, Jadoon Khan, M, Karsten, F, Groschner, LN, Waldeck-Weiermair, M, Rost, R, Hallström, S, Imamura, H, Graier, WF, Malli, R
    • Journal Title

      Molecular Biology of the Cell

      Volume: 25 Issue: 3 Pages: 368-379

    • DOI

      10.1091/mbc.e13-07-0433

    • Related Report
      2013 Annual Research Report 2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Glucose-stimulated single pancreatic islets sustain increased cytosolic ATP levels during initial Ca2+ influx and subsequent Ca2+ oscillations2013

    • Author(s)
      Tanaka T, Nagashima K, Inagaki N, Kioka H, Takashima S, Fukuoka H, Noji H, Kakizuka A and Imamura H
    • Journal Title

      J Biol Chem

      Volume: 289 Issue: 4 Pages: 2205-2216

    • DOI

      10.1074/jbc.m113.499111

    • NAID

      120005689188

    • Related Report
      2013 Annual Research Report 2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Evaluation of intra-mitochondrial ATP levels identifies G0/G1 switch gene 2 as a positive regulator of oxidative phosphorylation2013

    • Author(s)
      Kioka H, Kato H, Fujikawa M, Tsukamoto O, Suzuki T, Imamura H, Nakano A, Higo S, Yamazaki S, Matsuzaki T, Tkafuji K, Asanuma H, Asakura M, Minamino T, Shintani Y, Yoshida M, Noji H, Kitakaze M, Komuro I, Asano Y and Takashima S
    • Journal Title

      PNAS

      Volume: 111 Issue: 1 Pages: 273-278

    • DOI

      10.1073/pnas.1318547111

    • Related Report
      2013 Annual Research Report 2013 Final Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Genetically encoded fluorescent thermosensors visualize subcellular thermoregulation in living cells2013

    • Author(s)
      Shigeki Kiyonaka, Taketoshi Kajimoto, Reiko Sakaguchi, Daisuke Shinmi, Mariko Omatsu-Kanbe, Hiroshi Matsuura, Hiromi Imamura, Takenao Yoshizaki, Itaru Hamachi, Takashi Morii, Yasuo Mori
    • Journal Title

      Nature Methods

      Volume: 10 Issue: 12 Pages: 1232-1238

    • DOI

      10.1038/nmeth.2690

    • Related Report
      2013 Annual Research Report 2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] In vivo fluorescent ATP imaging of Drosophila melanogaster and Caenorhabditis elegans by using a genetically encoded fluorescent ATP biosensor optimized for low temperatures.2013

    • Author(s)
      Tsuyama, Taiichi; Kishikawa, Jun-ichi; Han, Yong-Woon; Harada, Yoshie; Tsubouchi, Asako; Noji, Hiroyuki; Kakizuka, Akira; Yokoyama, Ken; Uemura, Tadashi; Imamura, Hiromi.
    • Journal Title

      Analytical Chemistry

      Volume: 85(16) Issue: 16 Pages: 7889-7896

    • DOI

      10.1021/ac4015325

    • Related Report
      2013 Annual Research Report 2013 Final Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Role of PFKFB3-driven glycolysis in vessel sprouting2013

    • Author(s)
      De Bock, K, Georgiaudou, M, Schoors, S, Kuchnio, A, Rita Cantelmo, A, Wong, RW, Quaegebeur, A, Ghesquiére, B, Cauwehberghs, S, Eelen, G, Phng, LK, Betz, I, Tembuyser, B, Brepoels, K, Welti, J, Geudens, I, Segura, I, Cruys, B, Bifari, F, Decimo, I, Blanco, R, Wyns, S, Vangindertael, J, Rocha, S, Collins, R, Munck, S, Daelemans, D, Imamura, H, Devlieger, R, Rider, M, Van Veldhoven, PP, Schuit, F, Bartrons, R, Hofkens, J, Fraisi, P, Telang, S, DeBerardinis, RJ, Schoonjans, L, Vinckier, S, Chesney, J, Gerhardt, H, Dewerchin, M, Carmeliet, P
    • Journal Title

      Cell

      Volume: 154(3) Issue: 3 Pages: 651-63

    • DOI

      10.1016/j.cell.2013.06.037

    • Related Report
      2013 Annual Research Report 2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Distribution and Dynamics of Intracellular ATP Level, and their Regulation2013

    • Author(s)
      今村博臣、安東友美、藤川誠
    • Journal Title

      Seibutsu Butsuri

      Volume: 53 Issue: 1 Pages: 020-023

    • DOI

      10.2142/biophys.53.020

    • NAID

      10031139021

    • ISSN
      0582-4052, 1347-4219
    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Spatiotemporal correlations between cytosolic and mitochondrial Ca^<2+>signals using a novel red-shifted mitochondrial targeted cameleon2012

    • Author(s)
      Waldeck-Weiermair M, Alam MR, Khan MJ, Deak AT, Vishnu N, Karsten F, Imamura H, Graier WF, Malli R
    • Journal Title

      PLoS ONE

      Volume: 7 Issue: 9 Pages: e45917-e45917

    • DOI

      10.1371/journal.pone.0045917

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Hatsugai N, Koldenkova VP, Imamura H, Noji H, Nagai T2012

    • Author(s)
      Hatsugai N, Koldenkova VP, Imamura H, Noji H, Nagai T
    • Journal Title

      Plant. Cell Physiol

      Volume: 53 Issue: 10 Pages: 1768-1775

    • DOI

      10.1093/pcp/pcs119

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Assessing the actual contribution of IF1, an inhibitor of mitochondrial FoF1, to ATP homeostasis, cell growth, mitochondrial morphology and cell viability2012

    • Author(s)
      Fujikawa M, Imamura H, Nakamura J, Yoshida M
    • Journal Title

      J. Biol. Chem.

      Volume: 287 Issue: 22 Pages: 18781-18787

    • DOI

      10.1074/jbc.m112.345793

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Presentation] 膵臓ランゲルハンス島におけるATPとCa^<2+>の同時イメージング2014

    • Author(s)
      今村博臣
    • Organizer
      第3回発生におけるエネルギー代謝を考える会
    • Place of Presentation
      熊本大学発生医学研究所
    • Year and Date
      2014-02-13
    • Related Report
      2013 Final Research Report
  • [Presentation] 膵臓ランゲルハンス島におけるATPとCa^<2+>の同時イメージング2013

    • Author(s)
      田中喬、今村博臣
    • Organizer
      日本生体エネルギー研究会第39回討論会
    • Place of Presentation
      静岡県コンベンションアーツセンターグランシップ
    • Year and Date
      2013-12-19
    • Related Report
      2013 Final Research Report
  • [Presentation] ATPイメージングによる生細胞内ミトコンドリア機能の評価2013

    • Author(s)
      今村博臣
    • Organizer
      日本生化学会第86回年会シンポジウム「ミトコンドリアワールド : エネルギー生産から生体内環境保全まで」
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2013-09-11
    • Related Report
      2013 Final Research Report
  • [Presentation] 蛍光ATPバイオセンサーATeamを用いた膵島エネルギー代謝の解析2013

    • Author(s)
      今村博臣
    • Organizer
      第13回Islet Biology研究会
    • Place of Presentation
      日本都市センターホテル
    • Year and Date
      2013-07-20
    • Related Report
      2013 Final Research Report
  • [Presentation] 蛍光ATPバイオセンサーを用いたATP代謝の解析2013

    • Author(s)
      今村博臣
    • Organizer
      第50回日本臨床分子医学会学術集会第50回記念特別企画シンポジウム「分子イメージングの最前線」
    • Place of Presentation
      東京国際フォーラム
    • Year and Date
      2013-04-12
    • Related Report
      2013 Final Research Report
  • [Presentation] Imaging of intracellular ATP dynamics in single apoptotic cells2013

    • Author(s)
      Sakamoto S, Kakizuka A, Imamura H
    • Organizer
      11^<th> International Student Seminar
    • Place of Presentation
      Kyoto University, Japan
    • Year and Date
      2013-03-10
    • Related Report
      2013 Final Research Report
  • [Presentation] アポトーシスにおける細胞内ATPの挙動の解析2012

    • Author(s)
      坂本修一朗、野地博行、垣塚彰、今村博臣
    • Organizer
      日本生体エネルギー研究会第38回討論会
    • Place of Presentation
      岡山大学(岡山)
    • Year and Date
      2012-12-24
    • Related Report
      2013 Final Research Report
  • [Presentation] Fluorescent ATP biosensor optimized for imaging of cells from non-vertebrate model organisms2012

    • Author(s)
      Imamura H, Tsuyama T, Uemura T, Kakizuka A
    • Organizer
      International Symposia"New Frontiers of Metabolism Research in Biomedical Sciences"
    • Place of Presentation
      University of Tokyo, Japan
    • Year and Date
      2012-09-28
    • Related Report
      2013 Final Research Report
  • [Presentation] Genetically-encoded ATP biosensor for low temperatures2012

    • Author(s)
      Imamura H, Tsuyama T, Kishikawa J, Noji H, Kakizuka A, Yokoyama K, Uemura T
    • Organizer
      17^<th> European Bioenergetic Conference
    • Place of Presentation
      University of Freiburg, Germany
    • Year and Date
      2012-09-19
    • Related Report
      2013 Final Research Report
  • [Presentation] 蛍光ATPバイオセンサーを用いたATP代謝の解析

    • Author(s)
      今村博臣
    • Organizer
      第50回日本臨床分子医学会学術集会
    • Place of Presentation
      東京国際フォーラム
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] 蛍光ATPバイオセンサーATeamを用いた膵島エネルギー代謝の解析

    • Author(s)
      今村博臣
    • Organizer
      第13回Islet Biology 研究会
    • Place of Presentation
      日本都市センターホテル
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] ATPイメージングによる生細胞内ミトコンドリア機能の評価

    • Author(s)
      今村博臣
    • Organizer
      日本生化学会第86回年会
    • Place of Presentation
      パシフィコ横浜
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] 膵臓ランゲルハンス島におけるATPとCa2+の同時イメージング

    • Author(s)
      田中喬、今村博臣
    • Organizer
      日本生体エネルギー研究会 第39回討論会
    • Place of Presentation
      静岡県コンベンションアーツセンター
    • Related Report
      2013 Annual Research Report
  • [Presentation] 膵臓ランゲルハンス島におけるATPとCa2+の同時イメージング

    • Author(s)
      今村博臣
    • Organizer
      第3回 発生におけるエネルギー代謝を考える会
    • Place of Presentation
      熊本大学
    • Related Report
      2013 Annual Research Report
  • [Book] "ATP imaging in Xenopus laevis oocyte."In a book entitled "Sexual Reproduction in Animals and Plants"(Hitoshi Sawada, Naokazu Inoue, and Megumi Iwano)

    • Author(s)
      Ijiri, T.W., Kishikawa, J., Imamura, H., Iwao, Y., Yokoyama, K. and Sato K.
    • Publisher
      Springer Japan
    • Related Report
      2013 Final Research Report
  • [Remarks]

    • URL

      http://www.lif.kyoto-u.ac.jp/labs/imamura/index.html

    • Related Report
      2013 Final Research Report
  • [Remarks] インスリン分泌における重要因子が変動する様子を可視化

    • URL

      http://www.kyoto-u.ac.jp/ja/news_data/h/h1/news6/2013_1/140130_2.htm

    • Related Report
      2013 Annual Research Report
  • [Remarks] Imamura group HP

    • URL

      http://www.lif.kyoto-u.ac.jp/labs/imamura/Site/Top.html

    • Related Report
      2012 Research-status Report

URL: 

Published: 2013-05-31   Modified: 2019-07-29  

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