Mechanism analysis of redox imbalance caused by aberrant proteostasis and its recovery using fluorescence protein sensors visualizing redox states.

• Project/Area Number: 26292052
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Applied biochemistry
• Research Institution: Kyoto University
• Principal Investigator: Sakai Yasuyoshi 京都大学, (連合)農学研究科(研究院), 教授 (60202082)
• Co-Investigator(Kenkyū-buntansha): 寳関 淳 京都大学, 農学研究科, 特任准教授 (40423058)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥16,510,000 (Direct Cost: ¥12,700,000、Indirect Cost: ¥3,810,000); Fiscal Year 2016: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2015: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2014: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
• Keywords: レドックス / 蛍光タンパク質 / タンパク質代謝異常 / ケミカルバイオロジー

Outline of Final Research Achievements

Based on the redox sensors, roGFP and our developed Redoxfluor, we have created organelle-targeting super redox sensors and techniques monitoring intracellular local redox states in mammalian cells. Using these techniques, we have elucidated molecular mechanisms of abnormal redox states caused by aberrant intracellular protein metabolisms. Furthermore, we demonstrated food-derived compounds that recovered abnormal redox states as redox modulators.

Research Products

• [Journal Article] Synthesized Aβ42 Caused Intracellular Oxidative Damage, Leading to Cell Death, via Lysosome Rupture (2017)
  • Author(s): Yuki Oku, Kazuma Murakami, Kazuhiro Irie, Jun Hoseki, and Yasuyoshi Sakai
  • Journal Title: Cell Structure and Function Volume: 42 Issue: 1 Pages: 71-79
  • DOI: 10.1247/csf.17006
  • NAID: 130005665696
  • ISSN: 0386-7196, 1347-3700
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Development of a stable ERroGFP variant suitable for monitoring redox dynamics in the ER (2016)
  • Author(s): J. Hoseki, A. Oishi, T. Fujimura and Y. Sakai
  • Journal Title: Bioscience Reports Volume: 36 Issue: 2
  • DOI: 10.1042/bsr20160027
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Screening of dietary antioxidants against mitochondria-mediated oxidative stress by visualization of intracellular redox state (2016)
  • Author(s): S. Maharjan, Y. Sakai and J. Hoseki
  • Journal Title: Bioscience, Biotechonology and Biochemistry Volume: 80 Pages: 726-734
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Mitochondrial impairment triggers cytosolic oxidative stress and cell death following proteasome inhibition (2014)
  • Author(s): Sunita Maharjan, Masahide Oku, Masashi Tsuda, Jun Hoseki and Yasuyoshi Sakai
  • Journal Title: Scientific Reports Volume: 4 Issue: 1 Pages: 5896-5896
  • DOI: 10.1038/srep05896
  • NAID: 120005462594
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] 奥 勇紀、奥野 友紀子、萩原 正敏、寳関 淳、阪井 康能 (2017)
  • Author(s): 小胞体でのミスフォールドタンパク質蓄積に伴うレドックス異常回復剤の同定とその機序について
  • Organizer: 日本農芸化学会2017年大会
  • Place of Presentation: 京都女子大学 (京都市)
  • Year and Date: 2017-03-17
• [Presentation] メタノール資化性酵母の利用を出発点にした分子細胞生物学の発展 (2017)
  • Author(s): 阪井 康能
  • Organizer: 大阪大学工学研究科酵母リソース工学寄附講座 成果報告シンポジウム 「メタノール資化性酵母研究の展望」
  • Place of Presentation: 大阪大学 銀杏会館 (大阪府・吹田市)
  • Invited
• [Presentation] 植物の葉上におけるC1酵母の生存戦略とそれを支える分子細胞機能 (2017)
  • Author(s): 阪井 康能
  • Organizer: 第82回 酵母研究会
  • Place of Presentation: 北野工房のまち (神戸市)
  • Invited
• [Presentation] プロテアソーム阻害に伴う鉄代謝異常・酸化ストレスとミトコンドリア障害 (2016)
  • Author(s): 黒板 智博、Sunita Maharjan、寳関 淳、阪井 康能
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜 (横浜市)
  • Year and Date: 2016-11-30
• [Presentation] Gene-encoded Redox Probes, Application to Biology and Drug Screening (2016)
  • Author(s): Yasuyoshi Sakai
  • Organizer: FB3: Fluorescent Biomolecules and Their Building Blocks-Design and Applications
  • Place of Presentation: 天津大学 (天津、中国)
  • Year and Date: 2016-07-07
  • Int'l Joint Research / Invited
• [Presentation] プロテアソーム阻害による異常タンパク質がミトコンドリア障害を惹起する機構 (2016)
  • Author(s): 寳関 淳、Sunita Maharjan、阪井 康能
  • Organizer: 第68回日本細胞生物学会大会
  • Place of Presentation: 京都テルサ (京都市)
  • Year and Date: 2016-06-15
• [Presentation] 小胞体における異常タンパク質蓄積に伴うレドックス変化とこれを回復させる薬剤探索系の開発 (2016)
  • Author(s): 寳関 淳、奥 勇紀、藤村 多嘉朗、大石 麻水、奥野 友紀子、萩原 正敏、阪井 康能
  • Organizer: 第68回日本細胞生物学会大会
  • Place of Presentation: 京都テルサ (京都市)
  • Year and Date: 2016-06-15
• [Presentation] 細胞内の可視化を通して「わかったこと」「役立てること」 (2016)
  • Author(s): 阪井 康能
  • Organizer: 第10回中部大学ライフサイエンスフォーラム
  • Place of Presentation: 中部大学不言実行館アクティブホール（春日井市、愛知県）
  • Year and Date: 2016-01-13
  • Invited
• [Presentation] 生理的レドックス状態を可視化する分子プローブの開発と生理化学への応用 (2016)
  • Author(s): 阪井 康能
  • Organizer: 第37回「食と健康」研究会
  • Place of Presentation: サントリー ワールド リサーチセンター (京都府・精華町)
  • Invited
• [Presentation] 細胞内の局所的レドックスを指標にした生理化学 (2016)
  • Author(s): 阪井 康能
  • Organizer: 京都大学学際融合教育推進センター 生理化学研究ユニット 第６回公開シンポジウム 「Chemistryで紐解くPhysiology」
  • Place of Presentation: 京都大学農学研究科 (京都市)
  • Invited
• [Presentation] プロテアソーム阻害に伴う小胞体レドックス異常の機序 (2015)
  • Author(s): 藤村 多嘉朗、大石 麻水、Sunita Mahajan、寳関 淳、阪井 康能
  • Organizer: BMB2015(第38回日本分子生物学会年会、第88回日本生化学会大会 合同大会）
  • Place of Presentation: 神戸ポートアイランド(神戸市)
  • Year and Date: 2015-12-01
• [Presentation] 神経細胞内に蓄積したAβ42が細胞死を引き起こす機構 (2015)
  • Author(s): 奥 勇紀、村上 一馬、入江 一浩、寳関 淳、阪井 康能
  • Organizer: BMB2015(第38回日本分子生物学会年会、第88回日本生化学会大会 合同大会）
  • Place of Presentation: 神戸ポートアイランド(神戸市)
  • Year and Date: 2015-12-01
• [Presentation] The mechanisms underlying mitochondrial dysfunction under proteasome inhibition (2015)
  • Author(s): Sunita Maharjan、黒板 智博、寳関 淳、阪井 康能
  • Organizer: BMB2015(第38回日本分子生物学会年会、第88回日本生化学会大会 合同大会）
  • Place of Presentation: 神戸ポートアイランド(神戸市)
  • Year and Date: 2015-12-01
• [Presentation] The mechanism underlying mitochondrial dysfunction under proteasome inhibition (2015)
  • Author(s): S. Maharjan, J. Hoseki and Y. Sakai
  • Organizer: Cold Spring Harbor Asia Conferences-Mitochondria
  • Place of Presentation: Suzhou, China
  • Year and Date: 2015-10-12
  • Int'l Joint Research
• [Presentation] 蛍光可視化で探るオルガネラ ホメオスタシスの生理機能と障害 (2015)
  • Author(s): 阪井 康能
  • Organizer: 日本生化学会北海道支部共催講演会
  • Place of Presentation: 北海道大学地球環境科学研究院（札幌市）
  • Year and Date: 2015-09-08
  • Invited
• [Presentation] Workshop “Visualization of Local Redox State in Cells: Its Application to Biology and Drug Screening” (2015)
  • Author(s): Y. Sakai, M. Oku and J. Hoseki
  • Organizer: 15th International Conference on Oxidative Stress Reduction, Redox Homeostasis and Antioxidants 2015
  • Place of Presentation: Paris, France
  • Year and Date: 2015-06-22
  • Int'l Joint Research / Invited
• [Presentation] Symposium “How can we evaluate “physiological” redox state?” (2015)
  • Author(s): Y. Sakai
  • Organizer: 15th International Conference on Oxidative Stress Reduction, Redox Homeostasis and Antioxidants 2015
  • Place of Presentation: Paris, France
  • Year and Date: 2015-06-22
  • Int'l Joint Research / Invited
• [Presentation] Round Table Discussion “What are the Best & Adequate Methods to Evaluate Oxidative Stress & Antioxidants in vitro vs in vivo & in Humans?” (2015)
  • Author(s): Y. Sakai
  • Organizer: 15th International Conference on Oxidative Stress Reduction, Redox Homeostasis and Antioxidants 2015
  • Place of Presentation: Paris, France
  • Year and Date: 2015-06-22
  • Int'l Joint Research / Invited
• [Presentation] Intracellular Accumulation of Amyloid Beta Peptide Leads Lysosomal Membrane Permeabilization-Mediated Cell Death (2015)
  • Author(s): Y. Oku, K. Murakami, K. Irie, J. Hoseki and Y. Sakai
  • Organizer: 15th International Conference on Oxidative Stress Reduction, Redox Homeostasis and Antioxidants 2015
  • Place of Presentation: Paris, France
  • Year and Date: 2015-06-22
  • Int'l Joint Research
• [Presentation] Mitochondria trigger cytosolic oxidative stress under proteasome inhibition (2014)
  • Author(s): Sunita Maharjan, Masahide Oku, Jun Hoseki and Yasuyoshi Sakai
  • Organizer: Society for free radical biology and medicine's annual meeting 2014
  • Place of Presentation: Seatle (USA)
  • Year and Date: 2014-11-19 – 2014-11-23
• [Presentation] 異種タンパク質生産についてC1酵母を使って考える (2014)
  • Author(s): 阪井康能
  • Organizer: 新産業酵母研究会
  • Place of Presentation: 産総研：臨海副都心センター、東京
  • Year and Date: 2014-10-17
  • Invited
• [Presentation] Stable ERroGFP variants are suitable for detection of ER redox state (2014)
  • Author(s): Jun Hoseki, Asami Oishi and Yasuyoshi Sakai
  • Organizer: Gordon Research Conference, "Thiol-based redox regulation & signaling"
  • Place of Presentation: Girona (Spain)
  • Year and Date: 2014-07-20 – 2014-07-25
• [Presentation] 「遺伝子発現から薬効薬理評価～新リアルタイムイメージング～」、「異種タンパク質生産のTPOを考える：フォールディングとレドックス」 (2014)
  • Author(s): 阪井康能
  • Organizer: OYCバイオシンポジウム2014
  • Place of Presentation: 日本工業倶楽部会館、東京
  • Year and Date: 2014-07-04
  • Invited
• [Presentation] Antioxidants derived from foods protect redox aberration and cell death caused by proteasome inhibition (2014)
  • Author(s): Jun Hoseki
  • Organizer: Bordeaux-Kyoto
  • Place of Presentation: bordeaux (France)
  • Year and Date: 2014-05-05 – 2014-05-06
  • Invited
• [Remarks] タンパク質分解装置の活性低下が細胞死を引き起こす初期経路の同定と食品成分による回復
  • URL: http://www.kyoto-u.ac.jp/ja/research/research_results/2014/140731_1.html