Study of the mechanism of biosynthesis of polysulfurated proteins coupled with translation

2017 Fiscal Year Annual Research Report

• Project/Area Number: 16K15208
• Research Institution: Tohoku University
• Principal Investigator: 赤池 孝章 東北大学, 医学系研究科, 教授 (20231798)
• Co-Investigator(Kenkyū-buntansha): 藤井 重元 東北大学, 医学系研究科, 准教授 (00325333) ; 井田 智章 東北大学, 医学系研究科, 助教 (70570406)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: タンパク質ポリサルファ化 / システインポリスルフィド / システイン-tRNA合成酵素 / 活性イオウ分子種

Outline of Annual Research Achievements

本研究では、我々が最近発見したシステイン-tRNA合成酵素（CARS）による生物種横断的なシステインポリスルフィド生合成および翻訳に共役したタンパク質ポリサルファ化の分子メカニズムを解明し、その生理機能を明らかにすることを目的としている。昨年度までの本研究課題において、質量分析（LC-MS/MS）を用いた各種活性イオウ分子種のメタボローム解析およびタンパク質ポリサルファのゲルシフトアッセイとプロテオーム解析により、細菌および哺乳類のCARSのシステインパースルフィド合成活性とタンパク質ポリスルフィド化への関与を明らかにしてきた。本年度は、CARSによるシステインパースルフィド産生およびタンパク質ポリスルフィド化の分子機構について詳細に解析した。その結果、細菌および哺乳類の各種CARSのシステインパースルフィド合成活性の解析により、システインを基質としたシステインパースルフィド産生の酵素反応機構と活性発現に関与するアミノ酸残基を明らかにした。また、無細胞タンパク質合成系と各種アミノ酸変異CARSを用いた解析により、CARSは様々なタンパク質を普遍的にポリサルファ化することが明らかになった。さらに、ミトコンドリアに局在するCARS2を欠損させた細胞の解析により、CARS2によるタンパク質ポリサルファ化がミトコンドリアの伸長・分裂や膜電位形成などの機能制御に深く関わっていることが示された。CARSによる翻訳に共役したシステインパースルフィド産生とタンパク質ポリサルファ化は、ミトコンドリア機能をはじめとした細胞機能制御に重要な役割を果たしていることが示唆された。

Research Products

• [Int'l Joint Research] National Institute of Oncology(Hungary)
  • Country Name: Hungary
  • Counterpart Institution: National Institute of Oncology
• [Int'l Joint Research] University of Southampton(英国)
  • Country Name: UNITED KINGDOM
  • Counterpart Institution: University of Southampton
• [Int'l Joint Research] Sonoma State University/University of Vermont(米国)
  • Country Name: U.S.A.
  • Counterpart Institution: Sonoma State University/University of Vermont
• [Int'l Joint Research] German Cancer Research Center(Germany)
  • Country Name: Germany
  • Counterpart Institution: German Cancer Research Center
• [Journal Article] Involvement of nitric oxide/reactive oxygen species signaling via 8-nitro-cGMP formation in 1-methyl-4-phenylpyridinium ion-induced neurotoxicity in PC12 cells and rat cerebellar granule neurons (2018)
  • Author(s): Masuda K, Tsutsuki H, Kasamatsu S, Ida T, Takata T, Sugiura K, Nishida M, Watanabe Y, Sawa T, Akaike T, Ihara H.
  • Journal Title: Biochem Biophys Res Commun Volume: 495 Pages: 2165～2170
  • DOI: 10.1016/j.bbrc.2017.12.088
  • Peer Reviewed
• [Journal Article] 8-Nitro-cGMP is a promoter of osteoclast differentiation induced by RANKL (2018)
  • Author(s): Kaneko K, Miyamoto Y, Tsukuura R, Sasa K, Akaike T, Fujii S, Yoshimura K, Nagayama K, Hoshino M, Inoue S, Maki K, Baba K, Chikazu D, Kamijo R.
  • Journal Title: Nitric Oxide Volume: 72 Pages: 46～51
  • DOI: 10.1016/j.niox.2017.11.006
  • Peer Reviewed
• [Journal Article] Important role of endothelial caveolin-1 in the protective role of endothelium-dependent hyperpolarization against nitric oxide-mediated nitrative stress in microcirculation in mice (2018)
  • Author(s): Saito H, Godo S, Sato S, Ito A, Ikumi Y, Tanaka S, Ida T, Fujii S, Akaike T, Shimokawa H.
  • Journal Title: J Cardiovasc Pharmacol Volume: 71 Pages: 113-126
  • DOI: 10.1097/FJC.0000000000000552
  • Peer Reviewed
• [Journal Article] 8-Nitro-cGMP attenuates the interaction between SNARE complex and complexin through S-guanylation of SNAP-25 (2018)
  • Author(s): Kishimoto Y, Kunieda K, Kitamura A, Kakihana Y, Akaike T, Ihara H.
  • Journal Title: ACS Chem Neurosci Volume: 9 Pages: 217～223
  • DOI: 10.1021/acschemneuro.7b00363
  • Peer Reviewed
• [Journal Article] Cysteine perthiosulfenic acid (Cys-SSOH): A novel intermediate in thiol-based redox signaling? (2018)
  • Author(s): Heppner DE, Hristova M, Ida T, Mijuskovic A, Dustin CM, Bogdandi V, Fukuto JM, Dick TP, Nagy P, Li J, Akaike T, van der Vliet A.
  • Journal Title: Redox Biol Volume: 14 Pages: 379～385
  • DOI: 10.1016/j.redox.2017.10.006
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Persulfide synthases function coupled with translation and mediate sulfur respiration (2018)
  • Author(s): Fujii S, Sawa T, Motohashi H, Akaike T.
  • Journal Title: Br J Pharmacol Volume: - Pages: -
  • Peer Reviewed
• [Journal Article] 概論 レドックス疾患学：レドックス制御の破綻による病態と新たな疾患概念 (2018)
  • Author(s): 本橋ほづみ、赤池孝章、内田浩二、末松 誠.
  • Journal Title: 実験医学増刊「レドックス疾患学」 Volume: 36 Pages: 642～648
• [Journal Article] 硫黄呼吸の発見：ペルスルフィド産生酵素による新しいエネルギー代謝 (2018)
  • Author(s): 西村 明、赤池孝章.
  • Journal Title: 現代化学 Volume: 565 Pages: 55～59
• [Journal Article] Synthesis of L-cysteine derivatives containing stable sulfur isotopes and application of this synthesis to reactive sulfur metabolome (2017)
  • Author(s): Ono K, Jung M, Zhang T, Tsutsuki H, Sezaki H, Ihara H, Wei FY, Tomizawa K, Akaike T, Sawa T.
  • Journal Title: Free Radic Biol Med Volume: 106 Pages: 69～79
  • DOI: 10.1016/j.freeradbiomed.2017.02.023
  • Peer Reviewed
• [Journal Article] Quantitative determination of polysulfide in albumins, plasma proteins and biological fluid samples using a novel combined assays approach (2017)
  • Author(s): Ikeda M, Ishima Y, Shibata A, Chuang VTG, Sawa T, Ihara H, Watanabe H, Xian M, Ouchi Y, Shimizu T, Ando H, Ukawa M, Ishida T, Akaike T, Otagiri M, Maruyama T.
  • Journal Title: Anal Chim Acta Volume: 969 Pages: 18～25
  • DOI: 10.1016/j.aca.2017.03.027
  • Peer Reviewed
• [Journal Article] Reactive sulfur species inactivate Ca2+/calmodulin-dependent protein kinase IV via S-polysulfidation of its active-site cysteine residue (2017)
  • Author(s): Takata T, Ihara H, Hatano N, Tsuchiya Y, Akaike T, Watanabe Y.
  • Journal Title: Biochem J Volume: 474 Pages: 2547～2562
  • DOI: 10.1042/BCJ20170092
  • Peer Reviewed
• [Journal Article] Redox regulation of electrophilic signaling by reactive persulfides in cardiac cells (2017)
  • Author(s): Nishida M, Nishimura A, Matsunaga T, Motohashi H, Kasamatsu S, Akaike T.
  • Journal Title: Free Radic Biol Med Volume: 109 Pages: 132～140
  • DOI: 10.1016/j.freeradbiomed.2017.01.024
  • Peer Reviewed
• [Journal Article] Disruption of the structural and functional features of surfactant protein A by acrolein in cigarette smoke (2017)
  • Author(s): Takamiya R, Uchida K, Shibata T, Maeno T, Kato M, Yamaguchi Y, Ariki S, Hasegawa Y, Saito A, Miwa S, Takahashi H, Akaike T, Kuroki Y, Takahashi M.
  • Journal Title: Sci Rep Volume: 7 Pages: 8304
  • DOI: 10.1038/s41598-017-08588-5
  • Peer Reviewed / Open Access
• [Journal Article] 8-Nitro-cGMP promotes bone growth through expansion of growth plate cartilage (2017)
  • Author(s): Hoshino M, Kaneko K, Miyamoto Y, Yoshimura K, Suzuki D, Akaike T, Sawa T, Ida T, Fujii S, Ihara H, Tanaka J, Tsukuura R, Chikazu D, Mishima K, Baba K, Kamijo R.
  • Journal Title: Free Radic Biol Med Volume: 110 Pages: 63～71
  • DOI: 10.1016/j.freeradbiomed.2017.05.022
  • Peer Reviewed
• [Journal Article] Exposure to electrophiles impairs reactive persulfide-dependent redox signaling in neuronal cells (2017)
  • Author(s): Ihara H, Kasamatsu S, Kitamura A, Nishimura A, Tsutsuki H, Ida T, Ishizaki K, Toyama T, Yoshida E, Abdul Hamid H. Jung M, Matsunaga T, Fujii S, Sawa T, Nishida M, Kumagai Y, Akaike T.
  • Journal Title: Chem Res Toxicol Volume: 30 Pages: 1673～1684
  • DOI: 10.1021/acs.chemrestox.7b00120
  • Peer Reviewed
• [Journal Article] Chemical biology of hydropersulfides and related species: Possible roles in cellular protection and redox signaling (2017)
  • Author(s): Alvarez L, Bianco CL, Toscano JP, Lin J, Akaike T, Fukuto JM.
  • Journal Title: Antioxid Redox Signal Volume: 27 Pages: 622～633
  • DOI: 10.1089/ars.2017.7081
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Diversity and microevolution of CRISPR loci in Helicobacter cinaedi (2017)
  • Author(s): Tomida J, Morita Y, Shibayama K, Kikuchi K, Sawa T, Akaike T, Kawamura Y.
  • Journal Title: PLoS One Volume: 12 Pages: e0186241
  • DOI: 10.1371/journal.pone.0186241
  • Peer Reviewed / Open Access
• [Journal Article] Production of reactive persulfide species in chronic obstructive pulmonary disease (2017)
  • Author(s): Numakura T, Sugiura H, Akaike T, Ida T, Fujii S, Koarai A, Yamada M, Onodera K, Hashimoto Y, Tanaka R, Sato K, Shishikura Y, Hirano T, Yanagisawa S, Fujino N, Okazaki T, Tamada T, Hoshikawa Y, Okada Y, Ichinose M.
  • Journal Title: Thorax Volume: 72 Pages: 1074～1083
  • DOI: 10.1136/thoraxjnl-2016-209359
  • Peer Reviewed
• [Journal Article] Cysteinyl-tRNA synthetase governs cysteine polysulfidation and mitochondrial bioenergetics (2017)
  • Author(s): Akaike T, Ida T, Wei FY, Nishida M, Kumagai Y, Alam MM, Ihara H, Sawa T, Matsunaga T, Kasamatsu S, Nishimura A, Morita M, Tomizawa K, Nishimura A, Watanabe S, Inaba K, Shima H, Tanuma N, Jung M, Fujii S, Watanabe Y, Ohmuraya M, Nagy P, Feelisch M, Fukuto JM, Motohashi H.
  • Journal Title: Nat Commun Volume: 8 Pages: 1177
  • DOI: 10.1038/s41467-017-01311-y
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] ニトロ化環状ヌクレオチドと活性イオウ分子によるレドックスシグナル制御と生理機能 (2017)
  • Author(s): 藤井重元、赤池孝章.
  • Journal Title: 化学療法の領域 Volume: 33 Pages: 2194～2195
• [Journal Article] 活性イオウ分子：システインパースルフィドの新規生合成経路の発見とその生理機能 (2017)
  • Author(s): 西村 明、赤池孝章.
  • Journal Title: 硫酸と工業 Volume: 70 (11) Pages: 1～9
• [Presentation] Sulfur respiration in mammals and its implication for tumor biology (2018)
  • Author(s): Takaaki Akaike, Tomoaki Ida, Md. Morshedul Alam, Akira Nishimura, Masanobu Morita, Tetsuro Matsunaga, Hozumi Motohashi.
  • Organizer: The Cold Spring Harbor Asia Conference on Cancer & Metabolism
  • Int'l Joint Research / Invited
• [Presentation] システインパースルフィド産生酵素（Cysteine Persulfide Synthase, CPERS）の発見 (2017)
  • Author(s): 赤池孝章
  • Organizer: 第17回日本NO学会学術集会
  • Invited
• [Presentation] イオウ呼吸とイオウストレス：ほ乳類における新しいエネルギー代謝とレドックス病態 (2017)
  • Author(s): 赤池孝章、西村 明、井田智章、松永哲郎、守田匡伸、本橋ほづみ.
  • Organizer: 2017年度生理学研究所研究会「オルガネラダイナミクスの新規制御機構とその病態生理」
  • Invited
• [Presentation] イオウ呼吸とイオウストレス：新しいエネルギー代謝とレドックス病態の提案 (2017)
  • Author(s): 赤池孝章、西村 明、井田智章、松永哲郎、守田匡伸、藤井重元、本橋ほづみ.
  • Organizer: 第70回日本酸化ストレス学会学術集会
• [Presentation] Cysteine polysulfidation governed by cysteinyl-tRNA synthetase (CARSs) (2017)
  • Author(s): Takaaki Akaike
  • Organizer: Se2017 - 200 Years of Selenium Research (The 11th International Symposium on Selenium in Biology and Medicine and The 5th International Conference on Selenium in the Environment and Human Health)
  • Int'l Joint Research / Invited
• [Presentation] Cysteinyl-tRNA synthetase (CARS) controls endogenous hydropersulfide production and mediates mitochondrial functions and bioenegetics (2017)
  • Author(s): Takaaki Akaike
  • Organizer: S-Bio 2017 Plant and Human Sulfur Biology Conference 2017
  • Int'l Joint Research / Invited
• [Presentation] Discovery of a novel persulfide synthase that mediates mitochondrial sulfur respiration (2017)
  • Author(s): Takaaki Akaike
  • Organizer: 2nd Karolinska-Tohoku Joint Symposium on Medical Sciences
  • Int'l Joint Research / Invited
• [Presentation] Cysteinyl-tRNA synthetase governs endogenous hydropersulfide production and mediates mitochondrial functions and bioenergetics (2017)
  • Author(s): Takaaki Akaike
  • Organizer: AARS2017: IUBMB Focused Meeting on Aminoacyl-tRNA Synthetase
  • Int'l Joint Research
• [Presentation] Moonlighting functions of cysteinyl-tRNA synthetase: Cysteine hydropersulfide production and regulation of mitochondrial biogenesis and bioenergetics (2017)
  • Author(s): Takaaki Akaike, Akira Nishimura, Tomoaki Ida, Tetsuro Matsunaga, Masanobu Morita, Hozumi Motohashi.
  • Organizer: SfRBM 2017 - 24th Annual Meeting of the Society for Redox Biology and Medicine
  • Int'l Joint Research
• [Presentation] Cysteinyl-tRNA synthetase (CARS) は活性パースルフィド産生とミトコンドリア機能をコントロールしている (2017)
  • Author(s): 赤池孝章
  • Organizer: 2017年度生命科学系学会合同年次大会
  • Invited
• [Presentation] Cysteine persulfide formation via cysteinyl-tRNA synthetase and its involvement in mitochondrial bioenergetics (2017)
  • Author(s): Takaaki Akaike, Tomoaki Ida, Masanobu Morita, Shigemoto Fujii, Akira Nishimura, Tetsuro Matsunaga, Minkyung Jung, Hozumi Motohashi.
  • Organizer: 8th Joint Meeting of Society for Free Radical Research Australasia and Japan with International Symposium on Coenzyme Q10
  • Int'l Joint Research
• [Book] Nitric Oxide: Biology and Pathobiology. 3rd ed. (Ignarro LJ, Freeman BA, eds.) (2017)
  • Author(s): Sawa T, Kumagai Y, Akaike T.
  • Total Pages: 403
  • Publisher: Academic Press
  • ISBN: 978-0-12-804273-1
• [Remarks] 東北大学大学院医学系研究科 環境医学分野教室ホームページ
  • URL: http://www.toxicosci.med.tohoku.ac.jp