Quantitative analysis of ATP dynamics in photosynthetic organisms by a novel bioluminescence ATP biosensor

• Project/Area Number: 16K14709
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biophysics
• Research Institution: Kyoto University
• Principal Investigator: Imamura Hiromi 京都大学, 生命科学研究科, 准教授 (20422545)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 生物発光 / BRET / イメージング / 光合成 / ATP / ライブイメージング / エネルギー代謝

Outline of Final Research Achievements

We developed a novel genetically encoded BRET-based ATP biosensor, BTeam. It reacts specifically to ATP, and reports ATP concentrations in a range of 0.5 - 6.0 mM as BRET efficiencies. By measuring BRET efficiencies of BTeam-expressing cultured cells, we were able to quantitatively estimate ATP concentrations inside living cells. We also demonstrated that BTeam can report ATP concentrations inside a living organism using a zebrafish expressing BTeam. Currently, we are constructing a plant expressing BTeam in order to monitor ATP dynamics in plants during photosynthesis.

Research Products

• [Journal Article] General anesthetics cause mitochondrial dysfunction and reduction of intracellular ATP levels. (2018)
  • Author(s): Kishikawa JI, Inoue Y, Fujikawa M, Nishimura K, Nakanishi A, Tanabe T, Imamura H, Yokoyama K.
  • Journal Title: PLoS ONE Volume: 13 Issue: 1 Pages: e0190213-e0190213
  • DOI: 10.1371/journal.pone.0190213
  • Peer Reviewed / Open Access
• [Journal Article] Analysis of mitochondrial function in human induced pluripotent stem cells from patients with mitochondrial diabetes due to the A3243G mutation. (2018)
  • Author(s): Matsubara M, Kanda H, Imamura H, Inoue M, Noguchi M, Hosoda K, Kakizuka A, Nakao K.
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 949-949
  • DOI: 10.1038/s41598-018-19264-7
  • NAID: 120006502575
  • Peer Reviewed / Open Access
• [Journal Article] A Transient Rise in Free Mg 2+ Ions Released from ATP-Mg Hydrolysis Contributes to Mitotic Chromosome Condensation (2018)
  • Author(s): Maeshima Kazuhiro、Matsuda Tomoki、Shindo Yutaka、Imamura Hiromi、Tamura Sachiko、Imai Ryosuke、Kawakami Syoji、Nagashima Ryosuke、Soga Tomoyoshi、Noji Hiroyuki、Oka Kotaro、Nagai Takeharu
  • Journal Title: Current Biology Volume: 28 Issue: 3 Pages: 444-451
  • DOI: 10.1016/j.cub.2017.12.035
  • NAID: 120006380520
  • Peer Reviewed / Open Access
• [Journal Article] Human AK2 links intracellular bioenergetic redistribution to the fate of hematopoietic progenitors (2018)
  • Author(s): Oshima Koichi、Watanabe Akira、Nakahata Tatsutoshi、Saito Megumu K. et al.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 497 Issue: 2 Pages: 719-725
  • DOI: 10.1016/j.bbrc.2018.02.139
  • NAID: 120006466601
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Monitoring ATP dynamics in electrically active white matter tracts. (2017)
  • Author(s): Trevisiol A, Saab AS, Winkler U, Marx G, Imamura H, Moebius W, Kusch K, Nave KA, Hirrlinger J.
  • Journal Title: eLife Volume: 6
  • DOI: 10.7554/elife.24241
  • NAID: 120006318886
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cell competition with normal epithelial cells promotes apical extrusion of transformed cells through metabolic changes. (2017)
  • Author(s): Shunsuke Kon, et al.
  • Journal Title: Nature Cell Biology Volume: 印刷中 Issue: 5 Pages: 530-541
  • DOI: 10.1038/ncb3509
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] RLR-mediated antiviral innate immunity requires oxidative phosphorylation activity (2017)
  • Author(s): Yoshizumi T, Imamura H, Taku T, Kuroki T, Kawaguchi A, Ishikawa K, Nakada K, Koshiba T
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 5379-5379
  • DOI: 10.1038/s41598-017-05808-w
  • NAID: 120007128833
  • Peer Reviewed / Open Access
• [Journal Article] ATP Maintenance via Two Types of ATP Regulators Mitigates Pathological Phenotypes in Mouse Models of Parkinson's Disease (2017)
  • Author(s): Nakano Masaki、Imamura Hiromi、Sasaoka Norio、Yamamoto Masamichi、Uemura Norihito、Shudo Toshiyuki、Fuchigami Tomohiro、Takahashi Ryosuke、Kakizuka Akira
  • Journal Title: EBioMedicine Volume: 22 Pages: 225-241
  • DOI: 10.1016/j.ebiom.2017.07.024
  • NAID: 120006338063
  • Peer Reviewed / Open Access
• [Journal Article] Fusion protein analysis reveals the precise regulation between Hsp70 and Hsp100 during protein disaggregation. (2017)
  • Author(s): Hayashi S, Nakazaki Y, Kagii K, Imamura H, Watanabe YH.
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 8648-8648
  • DOI: 10.1038/s41598-017-08917-8
  • NAID: 120006364083
  • Peer Reviewed / Open Access
• [Journal Article] Visualization of long-term Mg2+ dynamics in apoptotic cells using a novel targetable fluorescent probe (2017)
  • Author(s): Matsui Y, Funato Y, Imamura H, Miki H, Mizukami S, Kikuchi K
  • Journal Title: Chem Sci. Volume: 8 Issue: 12 Pages: 8255-8264
  • DOI: 10.1039/c7sc03954a
  • Peer Reviewed / Open Access
• [Journal Article] Distinct intracellular Ca2+ dynamics regulate apical constriction and differentially contribute to neural tube closure. (2017)
  • Author(s): Suzuki, M., Sato, M., Koyama, H., Hara, Y., Hayashi, K., Yasue, N., Imamura, H., Fujimori, T., Nagai, T., Campbell, RE., Ueno, N.
  • Journal Title: Development. Volume: 144 Issue: 7 Pages: 1307-1316
  • DOI: 10.1242/dev.141952
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mitochondrial dysfunction induces dendritic loss via eIF2α phosphorylation. (2017)
  • Author(s): Taiichi Tsuyama, Asako Tsubouch, Tadao Usui, Hiromi Imamura, and Tadashi Uemura.
  • Journal Title: The Journal of Cell Biology Volume: 216(3) Issue: 3 Pages: 815-834
  • DOI: 10.1083/jcb.201604065
  • NAID: 120006360787
  • Peer Reviewed / Open Access
• [Journal Article] Application of FRET-Based Biosensor "ATeam" for Visualization of ATP Levels in the Mitochondrial Matrix of Living Mammalian Cells. (2017)
  • Author(s): Yoshida T, Alfaqaan S, Sasaoka N, Imamura H
  • Journal Title: Methods in Molecular Biology Volume: 1567 Pages: 231-243
  • DOI: 10.1007/978-1-4939-6824-4_14
  • ISBN: 9781493968220, 9781493968244
  • Peer Reviewed
• [Journal Article] BTeam, a Novel BRET-based Biosensor for the Accurate Quantification of ATP Concentration within Living Cells. (2016)
  • Author(s): Yoshida T, Kakizuka A, Imamura H.
  • Journal Title: Sci. Rep. Volume: 6 Issue: 1 Pages: 39618-39618
  • DOI: 10.1038/srep39618
  • NAID: 120005947215
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] アポトーシスにおける細胞内ATPダイナミクスのイメージング (2017)
  • Author(s): 今村博臣
  • Organizer: 第94回日本生理学会大会
  • Place of Presentation: アクトシティ浜松
  • Year and Date: 2017-03-29
  • Invited
• [Presentation] 分岐鎖アミノ酸に対するFRET型蛍光バイオセンサーの開発 (2017)
  • Author(s): 高橋世菜、垣塚彰、今村博臣
  • Organizer: 日本農芸化学会2017年度大会
  • Place of Presentation: 京都女子大学
  • Year and Date: 2017-03-19
• [Presentation] 蛍光バイオセンサーを用いた生細胞内可視化技術 (2017)
  • Author(s): 今村博臣
  • Organizer: 日本農芸化学会2017年度大会
  • Place of Presentation: 京都女子大学
  • Year and Date: 2017-03-18
  • Invited
• [Presentation] Imaging of intracellular ATP level revealed pannexin1-mediated programmed ATP decrease in apoptosis (2017)
  • Author(s): Imamura H, Sakamoto S, Yoshida T, Kakizuka A
  • Organizer: 19th International Union for Pure and Applied Biophysics Congress / 11th European Biophysics Congress
  • Int'l Joint Research
• [Presentation] FRET型ATPセンサーを用いたショウジョウバエ神経細胞におけるATPの濃度と消費速度の可視化 (2017)
  • Author(s): 津山泰一、坪内朝子、碓井理夫、今村博臣、上村匡
  • Organizer: 日本生体エネルギー研究会 第４３回討論会
• [Presentation] ルシフェラーゼ断片の相補的再構成を用いた新規発光ATPバイオセンサーの開発 (2017)
  • Author(s): 松岡篤志、垣塚彰、今村博臣
  • Organizer: 日本生体エネルギー研究会 第４３回討論会
• [Presentation] ツメガエル卵細胞におけるATPの役割の検証 (2017)
  • Author(s): 井尻貴之、岸川淳一、今村博臣、上野秀一、岩尾康宏、横山謙、佐藤賢一
  • Organizer: 日本生体エネルギー研究会 第４３回討論会
• [Presentation] FRET型蛍光バイオセンサーを用いたミトコンドリア分岐鎖アミノ酸輸送体の探索 (2017)
  • Author(s): 中嶋瞳、吉田有希、垣塚彰、今村博臣
  • Organizer: 日本生体エネルギー研究会 第４３回討論会
• [Presentation] 分岐鎖アミノ酸蛍光バイオセンサーの開発 (2017)
  • Author(s): 吉田有希、今村博臣
  • Organizer: 日本生体エネルギー研究会 第４３回討論会
• [Presentation] MPTPパーキンソン病モデルにおけるATP制御薬の発症緩和効果 (2017)
  • Author(s): 中野将希、今村博臣、笹岡紀男、山本正道、上村紀仁、首藤敏之、渕上智弘、高橋良輔、垣塚彰
  • Organizer: 日本生体エネルギー研究会 第４３回討論会
• [Presentation] 出芽酵母の細胞内ATP濃度変動を制御する仕組み (2017)
  • Author(s): 高稲正勝、今村博臣、吉田知史
  • Organizer: 日本生体エネルギー研究会 第４３回討論会
• [Presentation] 新規発光型ATPバイオセンサーの開発 (2016)
  • Author(s): 吉田有希、今村博臣
  • Organizer: 日本生体エネルギー研究会 第42回討論会
  • Place of Presentation: 名古屋工業大学
  • Year and Date: 2016-12-19
• [Presentation] ATP imaging revealed a mechanism of intracellular ATP changes during apoptosis (2016)
  • Author(s): 今村博臣
  • Organizer: 第54回日本生物物理学会年会
  • Place of Presentation: つくば国際会議場
  • Year and Date: 2016-11-27
• [Presentation] Molecular mechanism underlying intracellular ATP changes during apoptosis (2016)
  • Author(s): Imamura H, Sakamoto S, Yoshida T, Kakizuka A
  • Organizer: European Bioenergetics Conference 2016
  • Place of Presentation: Riva del Garda, Italy
  • Year and Date: 2016-07-02
  • Int'l Joint Research
• [Presentation] Development of a new BRET-based ATP indicator for quantitative intracellular ATP assay (2016)
  • Author(s): Yoshida T, Kakizuka A, Imamura H
  • Organizer: European Bioenergetics Conference 2016
  • Place of Presentation: Riva del Garda, Italy
  • Year and Date: 2016-07-02
  • Int'l Joint Research
• [Presentation] 生細胞ATPイメージングから酸素とATPの関係を探る (2016)
  • Author(s): 今村博臣
  • Organizer: 第16回日本蛋白質科学会年会
  • Place of Presentation: 福岡国際会議場
  • Year and Date: 2016-06-07
  • Invited