Reaction mechanism of a novel cysteine persulfide synthase and its function

• Project/Area Number: 20K07306
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 48040:Medical biochemistry-related
• Research Institution: Tohoku University
• Principal Investigator: Ida Tomoaki 東北大学, 医学系研究科, 助教 (70570406)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2022: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 活性イオウ分子種 / システインパースルフィド / システインtRNA合成酵素 / イオウメタボローム解析 / システインパースルフィド合成酵素 / イオウメタボローム / 活性イオウ / 活性イオウ分子

Outline of Research at the Start

システインパースルフィド(CysSSH）に代表される活性硫黄分子種は、生体内レドックスバランスを制御し、酸化ストレスに起因する様々な病態・疾患に対して防御的機能を発揮している。研究代表者らは、硫黄メタボロームを駆使して活性硫黄分子種の生体内生成を検証するなかで、cysteinyl-tRNA synthetase(CARS)が、主要なCysSSH合成酵素(CARS/cysteine persulfide synthase, CARS/CPERS)であることを見出した。本研究では、CysSSHの主要な産生酵素であるCARS/CPERSの酵素反応論と活性イオウ代謝制御機構を解明する。さらに、各種イオウ代謝関連遺伝子改変マウスを用いて、活性イオウ分子種の生体内生成動態と生理機能解析を行う。

Outline of Final Research Achievements

The enzymatic kinetics of CARS, the major enzyme producing cysteine persulfide (CysSSH), and its regulation mechanism of reactive sulfur metabolism were elucidated. Furthermore, the in vivo production levels of reactive sulfur species were shown using various sulfur metabolism-related gene-modified mice. In short, the CysSSH synthesis activity of CARS was found to be dependent on the microenvironment of the amino acid side chains around its KIIK motif. We also found that CARS has broad substrate specificity not only for cysteine but also for various thiol-containing compounds. This enzymatic reaction mechanism is suggested to be a unique reaction mechanism not involving classical Schiff bases. These findings suggest that CARS is involved in redox regulation by producing various persulfides.

Academic Significance and Societal Importance of the Research Achievements

これまで、CARSによるCysSSH合成機構は不明であった。本研究により、CARSによる各種パースルフィド合成機構が明らかになった。これにより、CARSによるCysSSH合成活性は、これまで全く知られていないCARSのユニークなmoonlighting functionであり、その酵素反応論の解明はレドックス研究の進展に大きく貢献した。さらに、活性イオウ分子種の生理学的役割の解明に大きく貢献し、予防医学・創薬などのトランスレーショナルな展開が期待された。

Research Products

• [Int'l Joint Research] York University(カナダ)
• [Int'l Joint Research] Massachusetts General Hospital(米国)
• [Int'l Joint Research] National Institute of Oncology(ハンガリー)
• [Int'l Joint Research] National Institute of Oncology(ハンガリー)
• [Journal Article] Synthesis of sulfides and persulfides is not impeded by disruption of three canonical enzymes in sulfur metabolism (2023)
  • Author(s): Zainol Abidin Qamarul Hafiz、Ida Tomoaki、Morita Masanobu、Matsunaga Tetsuro、Nishimura Akira、Jung Minkyung、Hassan Naim、Takata Tsuyoshi、Ishii Isao、Kruger Warren、Wang Rui、Motohashi Hozumi、Tsutsui Masato、Akaike Takaaki
  • Journal Title: Antioxidants Volume: 12 Issue: 4 Pages: 868-868
  • DOI: 10.3390/antiox12040868
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Polysulfide metabolizing enzymes influence SqrR-mediated sulfide-induced transcription by impacting intracellular polysulfide dynamics (2023)
  • Author(s): Shimizu Takayuki、Ida Tomoaki、Antelo Giuliano T、Ihara Yuta、Fakhoury Joseph N、Masuda Shinji、Giedroc David P、Akaike Takaaki、Capdevila Daiana A、Masuda Tatsuru
  • Journal Title: PNAS Nexus Volume: 2 Issue: 3 Pages: 1-11
  • DOI: 10.1093/pnasnexus/pgad048
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cysteine hydropersulfide reduces lipid peroxidation and protects against myocardial ischaemia-reperfusion injury - Are endogenous persulfides mediators of ischaemic preconditioning? (2023)
  • Author(s): Griffiths Kayleigh、Ida Tomoaki、Morita Masanobu、Lamb Reece J.、Lee Jordan J.、Frenneaux Michael P.、Fukuto Jon M.、Akaike Takaaki、Feelisch Martin、Madhani Melanie
  • Journal Title: Redox Biology Volume: 60 Pages: 102605-102605
  • DOI: 10.1016/j.redox.2023.102605
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Regulation of nitric oxide/reactive oxygen species redox signaling by nNOS splicing variants. (2022)
  • Author(s): Kasamatsu Shingo, Tsutsuki Hiroyasu, Ida Tomoaki, Sawa Tomohiro, Watanabe Yasuo, Akaike Takaaki, Ihara Hideshi.
  • Journal Title: Nitric Oxide Volume: 120 Pages: 44-52
  • DOI: 10.1016/j.niox.2022.01.004
  • Peer Reviewed
• [Journal Article] 8-Nitro-cGMP suppresses mineralization by mouse osteoblasts (2022)
  • Author(s): Kaneko K, Miyamoto Y, Ida T, Morita M, Yoshimura K, Nagasaki K, Toba K, Sugisaki R, Motohashi H, Akaike T, Chikazu D, Kamijo R
  • Journal Title: Journal of Clinical Biochemistry and Nutrition Volume: 71 Issue: 3 Pages: 191-197
  • DOI: 10.3164/jcbn.21-129
  • ISSN: 0912-0009, 1880-5086
  • Peer Reviewed / Open Access
• [Journal Article] GRIM-19 is a target of mycobacterial Zn2+ metalloprotease 1 and indispensable for NLRP3 inflammasome activation. (2022)
  • Author(s): Kurane, T., Matsunaga, T., Ida, T., Sawada, K., Nishimura, A., Fukui, M., Umemura, M., Nakayama, M., Ohara, N., Matsumoto, S., Akaike, T., Matsuzaki, G., Takaesu, G.
  • Journal Title: FASEB Journal Volume: 36 Issue: 1
  • DOI: 10.1096/fj.202101074rr
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Methods in sulfide and persulfide research. (2021)
  • Author(s): Takata Tsuyoshi, Jung Minkyung, Matsunaga Tetsuro, Ida Tomoaki, Morita Masanobu, Motohashi Hozumi, Shen Xinggui, Kevil Christopher G., Fukuto Jon M., Akaike Takaaki.
  • Journal Title: Nitric Oxide Volume: 116 Pages: 47-64
  • DOI: 10.1016/j.niox.2021.09.002
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Reprogrammed transsulfuration promotes basal-like breast tumor progression via realigning cellular cysteine persulfidation. (2021)
  • Author(s): Erdelyi Katalin, Ditroi Tamas, Johansson Henrik J., Czikora Agnes, Balog Noemi, Silwal-Pandit Laxmi, Ida Tomoaki, Olasz Judit, Hajdu Dorottya, Matrai Zoltan, Csuka Orsolya, Uchida Koji, Tovari Jozsef, Engebraten Olav, Akaike Takaaki, Borresen Dale Anne-Lise, Kasler Miklos, Lehtio Janne, Nagy Peter.
  • Journal Title: Proceedings of the National Academy of Sciences of the United States of America Volume: 118 Issue: 45
  • DOI: 10.1073/pnas.2100050118
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Sulfide catabolism ameliorates hypoxic brain injury. (2021)
  • Author(s): Marutani E, Morita M, Hirai S, Kai S, Grange RMH, Miyazaki Y, Nagashima F, Traeger L, Magliocca A, Ida T, Matsunaga T, Batten A, Li R, Tanaka T, Ikeda T, Nakagawa A, Atochin DN, Ihara H, Olenchock BA, Shen X, Nishida M, Hanaoka K, Kevil CG, Xian M, Bloch DB, Akaike T, Hindle AG, Motohashi H, Ichinose F, et al.
  • Journal Title: Nature Communications Volume: 12 Issue: 1 Pages: 3108-3108
  • DOI: 10.1038/s41467-021-23363-x
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] On-tissue polysulfide visualization by surface-enhanced Raman spectroscopy benefits patients with ovarian cancer to predict post-operative chemosensitivity (2021)
  • Author(s): Honda Kazufumi、Hishiki Takako、Yamamoto Sohe、Naito Yoshiko、Matsuura Tomomi、Iwaisako Keiko、Masui Toshihiko、Sakuma Tomohiro、Matsubara Osamu、Huang Wilber、Ida Tomoaki、Akaike Takaaki、Masugi Yohei、Sakamoto Michiie、Kato Tomoyasu、Kabe Yasuaki、Suematsu Makoto
  • Journal Title: Redox Biology Volume: 41 Pages: 101926-101926
  • DOI: 10.1016/j.redox.2021.101926
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] High-Precision Sulfur Metabolomics Innovated by a New Specific Probe for Trapping Reactive Sulfur Species (2021)
  • Author(s): Kasamatsu Shingo、Ida Tomoaki、Koga Taisei、Asada Kosho、Motohashi Hozumi、Ihara Hideshi、Akaike Takaaki
  • Journal Title: Antioxidants & Redox Signaling Volume: in press Issue: 18 Pages: 1407-1419
  • DOI: 10.1089/ars.2020.8073
  • Peer Reviewed / Open Access
• [Journal Article] Loss of cell wall integrity genes cpxA and mrcB causes flocculation in Escherichia coli (2021)
  • Author(s): Sugawara Keita、Toyoda Hayato、Kimura Mami、Hayasaka Shunsuke、Saito Hiromi、Kobayashi Hiroshi、Ihara Kunio、Ida Tomoaki、Akaike Takaaki、Ando Eiji、Hyodo Mamoru、Hayakawa Yoshihiro、Hamamoto Shin、Uozumi Nobuyuki
  • Journal Title: Biochemical Journal Volume: 478 Issue: 1 Pages: 41-59
  • DOI: 10.1042/bcj20200723
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Measuring Reactive Sulfur Species and Thiol Oxidation States: Challenges and Cautions in Relation to Alkylation-Based Protocols (2020)
  • Author(s): Nagy Peter、Doka Eva、Ida Tomoaki、Akaike Takaaki
  • Journal Title: Antioxidants & Redox Signaling Volume: 33 Issue: 16 Pages: 1174-1189
  • DOI: 10.1089/ars.2020.8077
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Glutathione Trisulfide Prevents Lipopolysaccharide-induced Inflammatory Gene Expression in Retinal Pigment Epithelial Cells (2020)
  • Author(s): Tawarayama Hiroshi、Suzuki Noriyuki、Inoue-Yanagimachi Maki、Himori Noriko、Tsuda Satoru、Sato Kota、Ida Tomoaki、Akaike Takaaki、Kunikata Hiroshi、Nakazawa Toru
  • Journal Title: Ocular Immunology and Inflammation Volume: in press Issue: 4 Pages: 1-12
  • DOI: 10.1080/09273948.2020.1833224
  • Peer Reviewed
• [Journal Article] 8-Nitro-cGMP modulates exocytosis in adrenal chromaffin cells (2020)
  • Author(s): Tsutsuki Hiroyasu、Kasamatsu Shingo、Kunieda Kohei、Ida Tomoaki、Sawa Tomohiro、Sasakawa Nobuyuki、Akaike Takaaki、Ihara Hideshi
  • Journal Title: Biochemical and Biophysical Research Communications Volume: in press Issue: 1 Pages: 225-230
  • DOI: 10.1016/j.bbrc.2020.03.045
  • Peer Reviewed / Open Access
• [Presentation] 細菌から動物まで種横断的に保存された超硫黄分子合成経路の発見 (2023)
  • Author(s): 井田智章、ジョンミンキョン、松永哲郎、守田匡伸、緒方星陵、高田 剛、海野雄加、本橋ほづみ、赤池孝章
  • Organizer: 第96回日本細菌学会総会
• [Presentation] Discovery of new persulfide synthases that are expressed among all organisms during their evolution (2022)
  • Author(s): Tomoaki Ida, Minkyung Jung, Seiryo Ogata, Masanobu Morita, Tetsuro Matsunaga, Yuka Unno, Tsuyoshi Takata, Ayumu Kobayashi, Yasuhiro Ishimaru, Nobuyuki Uozumi, Hozumi Motohashi, Takaaki Akaike
  • Organizer: The 12th International Conference on the Biology, Chemistry, and Therapeutic Applications of Nitric Oxide
  • Int'l Joint Research
• [Presentation] アミノアシル-tRNA合成酵素による種横断的超硫黄分子合成機構の解明 (2022)
  • Author(s): 井田智章、Jung Minkyung、松永哲郎、守田匡伸、高田 剛、本橋ほづみ、赤池孝章
  • Organizer: 第75回日本酸化ストレス学会学術集会
• [Presentation] 呼気オミックスによる生体防御応答解析 (2022)
  • Author(s): 井田智章、高田 剛、松永哲郎、守田匡伸、Jung Minkyung、 本橋ほづみ、赤池孝章
  • Organizer: 第33回日本生体防御学会学術総会
• [Presentation] 呼気オミックスから空間オミックス解析への展開 (2022)
  • Author(s): 井田智章、緒方星陵、松永哲郎、守田匡伸、高田 剛、海野雄加、Jung Minkyung、安西 眸、高奈秀匡、太田 信、本橋ほづみ、赤池孝章
  • Organizer: 第22回分子予防環境医学研究会
• [Presentation] 細菌におけるアミノアシル-tRNA合成酵素によるシステインパースルフィド合成機構の発見 (2022)
  • Author(s): 井田智章、ジョンミンキョン、西村 明、松永哲郎、守田匡伸、高田 剛、本橋ほづみ、赤池孝章
  • Organizer: 第95回日本細菌学会総会
• [Presentation] 呼気オミックスと病原体モニタリング (2022)
  • Author(s): 井田智章、赤池孝章
  • Organizer: 第95回日本細菌学会総会
  • Invited
• [Presentation] システインパースルフィド合成酵素の酵素反応機構とその機能解析 (2021)
  • Author(s): 井田智章、守田匡伸、松永哲郎、Jung Minkyung、高田 剛、本橋ほづみ、赤池孝章
  • Organizer: 第74回日本酸化ストレス学会 第21回日本NO学会 合同学術集会
• [Presentation] 超硫黄代謝物解析と呼気オミックスへの展開 (2021)
  • Author(s): 井田智章、松永哲郎、高田 剛、守田匡伸、Jung Minkyung、 澤 智裕、本橋ほづみ、赤池孝章
  • Organizer: 第94回日本生化学会大会
• [Presentation] 超硫黄分子による生体防御と超硫黄オミックス開発 (2021)
  • Author(s): 井田智章、松永哲郎、高田 剛、守田匡伸、Jung Minkyung、 澤 智裕、本橋ほづみ、赤池孝章
  • Organizer: 第33回日本生体防御学会学術総会
• [Presentation] 超硫黄オミックスの開発に向けた基盤研究 (2021)
  • Author(s): 井田智章、松永哲郎、高田 剛、守田匡伸、Jung Minkyung、 澤 智裕、本橋ほづみ、赤池孝章
  • Organizer: 生理研研究会2021 生命を支える硫黄生物学の最前線
• [Presentation] アミノアシル-tRNA合成酵素による超硫黄分子合成機構の解明 (2021)
  • Author(s): 井田智章、ジョンミンキョン、松永哲郎、守田匡伸、高田 剛、 本橋ほづみ、赤池孝章
  • Organizer: 第21回分子予防環境医学研究会大会
• [Presentation] NADPH oxidaseによる活性硫黄代謝リモデリングと感染防御機構 (2020)
  • Author(s): 井田智章、松永哲郎、守田匡伸、高田 剛、本橋ほづみ、住本英樹、赤池孝章
  • Organizer: 第93回日本細菌学会総会
• [Presentation] 新規システインパースルフィド合成酵素の同定と酵素反応機構の解明 (2020)
  • Author(s): 井田智章、西村 明、守田匡伸、Jung Minkyung、松永哲郎、居原 秀、 本橋ほづみ、赤池孝章
  • Organizer: 第73回日本酸化ストレス学会 第20回日本NO学会 合同学術集会
• [Presentation] 新型コロナウイルスオミックス解析システムの開発 (2020)
  • Author(s): 井田智章、高田 剛、守田匡伸、松永哲郎、Jung Minkyung、 澤 智裕、杉浦久敏、本橋ほづみ、赤池孝章
  • Organizer: 第31回日本生体防御学会学術総会
• [Remarks] 東北大学大学院医学系研究科環境医学分野HP
  • URL: http://www.toxicosci.med.tohoku.ac.jp/index.
• [Remarks] 東北大学大学院医学系研究科環境医学分野HP
  • URL: http://www.toxicosci.med.tohoku.ac.jp/index.html