Elucidation of ryanodine receptor channel gating mechanism at atomic level

• Project/Area Number: 16K08507
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General physiology
• Research Institution: Juntendo University
• Principal Investigator: Murayama Takashi 順天堂大学, 医学部, 准教授 (10230012)
• Project Period (FY): 2016-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: リアノジン受容体 / カルシウムイオン / イオンチャネル / 動作原理 / 骨格筋 / 心筋 / カルシウムチャネル / 構造活性相関 / 筋疾患 / カフェイン / 筋小胞体 / カルシウム / 疾患変異 / 作動機構 / 構造モデル / チャネル / 構造解析

Outline of Final Research Achievements

To understand the gating mechanism of ryanodine receptor (RyR), a calcium-release channel of sarcoplasmic reticulum, we combined functional analysis of RyR carrying disease-associated mutations with its structural model at near atomic resolution. We have elucidated for the first time the molecular mechanism of action of caffeine, which has long been known as a drug that increases Ca2+ sensitivity. In addition, we identified a series of interaction sites from the cytoplasmic region to the transmembrane region that require channel opening by Ca2+ binding.

Academic Significance and Societal Importance of the Research Achievements

本研究では変異体の機能解析と高分解能構造モデルを組み合わせることで、RyRの動作原理の本質に原子レベルで迫ることができた。特にカフェインによるCa2+感受性増大機構の解明は世界初であり、RyR制御機構における重要な成果である。本研究成果はリガンド作動性チャネルの動作原理として、イオンチャネル研究における大きな一歩である。また、本手法はRyRに特徴的な巨大な細胞質領域にも応用が可能である。細胞質領域には疾患変異が集中しているため、今後の研究により疾患変異の分子機構や薬物開発が大きく加速することが期待される。

Research Products

• [Int'l Joint Research] Virginia Commonwealth University(米国)
• [Int'l Joint Research] University of Leeds(英国)
• [Int'l Joint Research] Virginia Commonwealth University(米国)
• [Journal Article] Co-Phenotype of Left Ventricular Non-Compaction Cardiomyopathy and Atypical Catecholaminergic Polymorphic Ventricular Tachycardia in Association With R169Q, a <i>Ryanodine Receptor Type 2</i> Missense Mutation (2020)
  • Author(s): Nozaki Y, Kato Y, Uike K, Yamamura K, Kikuchi M, Yasuda M, Ohno S, Horie M, Murayama T, Kurebayashi N, Horigome H
  • Journal Title: Circulation Journal Volume: 84 Issue: 2 Pages: 226-234
  • DOI: 10.1253/circj.CJ-19-0720
  • NAID: 130007789920
  • ISSN: 1346-9843, 1347-4820
  • Year and Date: 2020-01-24
  • Peer Reviewed / Open Access
• [Journal Article] TRIC-A Channel Maintains Store Calcium Handling by Interacting With Type 2 Ryanodine Receptor in Cardiac Muscle (2020)
  • Author(s): Zhou X, Park KH, Yamazaki D, Lin PH, Nishi M, Ma Z, Qiu L, Murayama T, Zou X, Takeshima H, Zhou J, Ma J.
  • Journal Title: Circulation Reserach Volume: 126 Issue: 4 Pages: 417-435
  • DOI: 10.1161/circresaha.119.316241
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Insights into channel modulation mechanism of RYR1 mutants using Ca2+ imaging and molecular dynamics (2020)
  • Author(s): Yamazawa Toshiko、Ogawa Haruo、Murayama Takashi、Yamaguchi Maki、Oyamada Hideto、Suzuki Junji、Kurebayashi Nagomi、Kanemaru Kazunori、Oguchi Katsuji、Sakurai Takashi、Iino Masamitsu
  • Journal Title: The Journal of General Physiology Volume: 152 Issue: 1 Pages: 0-0
  • DOI: 10.1085/jgp.201812235
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Assays for Modulators of Ryanodine Receptor (RyR)/Ca 2+ Release Channel Activity for Drug Discovery for Skeletal Muscle and Heart Diseases (2019)
  • Author(s): Murayama Takashi、Kurebayashi Nagomi
  • Journal Title: Current Protocols in Pharmacology Volume: 87 Issue: 1 Pages: 0-0
  • DOI: 10.1002/cpph.71
  • Peer Reviewed / Open Access
• [Journal Article] Structural development of a type-1 ryanodine receptor (RyR1) Ca2+-release channel inhibitor guided by endoplasmic reticulum Ca2+ assay (2019)
  • Author(s): Mori Shuichi、Iinuma Hiroto、Manaka Noriaki、Ishigami-Yuasa Mari、Murayama Takashi、Nishijima Yoshiaki、Sakurai Akiko、Arai Ryota、Kurebayashi Nagomi、Sakurai Takashi、Kagechika Hiroyuki
  • Journal Title: European Journal of Medicinal Chemistry Volume: 179 Pages: 837-848
  • DOI: 10.1016/j.ejmech.2019.06.076
  • Peer Reviewed
• [Journal Article] Conversion of Ca2+ oscillation into propagative electrical signals by Ca2+-activated ion channels and connexin as a reconstituted Ca2+ clock model for the pacemaker activity (2019)
  • Author(s): Saeki Takanori、Kimura Taisuke、Hashidume Keigo、Murayama Takashi、Yamamura Hisao、Ohya Susumu、Suzuki Yoshiaki、Nakayama Shinsuke、Imaizumi Yuji
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 510 Issue: 2 Pages: 242-247
  • DOI: 10.1016/j.bbrc.2019.01.080
  • Peer Reviewed / Open Access
• [Journal Article] Dysfunction of muscle contraction with impaired intracellular Ca2+ handling in skeletal muscle and the effect of exercise training in male db/db mice (2019)
  • Author(s): Eshima Hiroaki、Tamura Yoshifumi、Kakehi Saori、Nakamura Kyoko、Kurebayashi Nagomi、Murayama Takashi、Kakigi Ryo、Sakurai Takashi、Kawamori Ryuzo、Watada Hirotaka
  • Journal Title: Journal of Applied Physiology Volume: 126 Issue: 1 Pages: 170-182
  • DOI: 10.1152/japplphysiol.00048.2018
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A tryptophan residue in the caffeine-binding site of the ryanodine receptor regulates Ca2+ sensitivity (2018)
  • Author(s): Murayama Takashi、Ogawa Haruo、Kurebayashi Nagomi、Ohno Seiko、Horie Minoru、Sakurai Takashi
  • Journal Title: Communications Biology Volume: 1 Issue: 1 Pages: 98-98
  • DOI: 10.1038/s42003-018-0103-x
  • Peer Reviewed / Open Access
• [Journal Article] Efficient high-throughput screening by ER Ca2+measurement to identify inhibitors of ryanodine receptor Ca2+-release channels (2018)
  • Author(s): Murayama Takashi、Kurebayashi Nagomi、Ikegami-Yuasa Mari、Mori Shuichi、Suzuki Yukina、Akima Ryunosuke、Ogawa Haruo、Suzuki Junji、Kanemaru Kazunori、Oyamada Hideto、Kiuchi Yuji、Iino Masamitsu、Kagechika Hiroyuki、Sakurai Takashi
  • Journal Title: Molecular Pharmacology Volume: - Issue: 1 Pages: 722-730
  • DOI: 10.1124/mol.117.111468
  • Peer Reviewed / Open Access
• [Journal Article] Fam64a is a novel cell cycle promoter of hypoxic fetal cardiomyocytes in mice (2017)
  • Author(s): Hashimoto Ken、Kodama Aya、Honda Takeshi、Hanashima Akira、Ujihara Yoshihiro、Murayama Takashi、Nishimatsu Shin-ichiro、Mohri Satoshi
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 4486-4486
  • DOI: 10.1038/s41598-017-04823-1
  • Peer Reviewed / Open Access
• [Journal Article] Dynactin has two antagonistic regulatory domains and exerts opposing effects on dynein motility (2017)
  • Author(s): Kobayashi Takuya、Miyashita Takuya、Murayama Takashi、Toyoshima Yoko Y.
  • Journal Title: PLOS ONE Volume: 12 Issue: 8 Pages: e0183672-e0183672
  • DOI: 10.1371/journal.pone.0183672
  • Peer Reviewed / Open Access
• [Journal Article] Long-term, but not short-term high-fat diet induces fiber composition changes and impaired contractile force in mouse fast-twitch skeletal muscle (2017)
  • Author(s): Eshima H,Tamura Y, Kakehi S, Kurebayashi N, Murayama T, Nakamura K, Kakigi R,Okada T,Sakurai T, Kawamori R, Watada H
  • Journal Title: Physiological reports Volume: 5 Issue: 7 Pages: e13250-e13250
  • DOI: 10.14814/phy2.13250
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Therapeutic potential of ghrelin and des-acyl ghrelin against chemotherapy-induced cardiotoxicity (2017)
  • Author(s): Nonaka Miki、Kurebayashi Nagomi、Murayama Takashi、Sugihara Masami、Terawaki Kiyoshi、Shiraishi Seiji、Miyano Kanako、Hosoda Hiroshi、Kishida Shosei、Kangawa Kenji、Sakurai Takashi、Uezono Yasuhito
  • Journal Title: Endocrine Journal Volume: 64 Issue: Suppl. Pages: S35-S39
  • DOI: 10.1507/endocrj.64.S35
  • NAID: 130007307943
  • ISSN: 0918-8959, 1348-4540
  • Peer Reviewed / Open Access
• [Journal Article] Extensive Ca2+ leak through K4750Q cardiac ryanodine receptors caused by cytosolic and luminal Ca2+ hypersensitivity. (2016)
  • Author(s): Uehara A, Horie M, et al.
  • Journal Title: Journal of General Physiology Volume: 149 Issue: 2 Pages: 199-218
  • DOI: 10.1085/jgp.201611624
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A type 2 ryanodine receptor variant associated with reduced Ca2+ release and short-coupled torsade de pointe ventricular arrhythmia. (2016)
  • Author(s): Fujii Y, Horie M, et al.
  • Journal Title: Heart Rhythm Volume: 14 Issue: 1 Pages: 98-107
  • DOI: 10.1016/j.hrthm.2016.10.015
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Genotype-phenotype correlations of malignant hyperthermia and central core disease mutations in the central region of the RYR1 channel. (2016)
  • Author(s): Murayama, T., Kurebayashi, N., Ogawa, H., Yamazawa, T., Oyamada, H., Suzuki, J., Kanemaru, K., Oguchi, K., Iino, M. and Sakurai, T.
  • Journal Title: Hum Mutat. Volume: 37 Issue: 11 Pages: 1231-1241
  • DOI: 10.1002/humu.23072
  • Peer Reviewed / Open Access
• [Journal Article] Ultrastructural analysis of self-associated RyR2s. (2016)
  • Author(s): Cabra V, Murayama T, Samso M
  • Journal Title: Biophysical Journal Volume: 110 Issue: 12 Pages: 2651-2662
  • DOI: 10.1016/j.bpj.2016.05.013
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Nitric oxide-induced activation of the type 1 ryanodine receptor is critical for epileptic seizure-induced neuronal cell death. (2016)
  • Author(s): Mikami, Y., Kanemaru, K., Okubo, Y., Nakaune, T., Suzuki, J., Shibata, K., Sugiyama, H., Koyama, R., Murayama, T., Ito, A., Yamazawa, T., Ikegaya, Y., Sakurai, T., Saito, N., Kakizawa, S. and Iino, M.
  • Journal Title: EBioMedicine. Volume: 11 Pages: 253-261
  • DOI: 10.1016/j.ebiom.2016.08.020
  • Peer Reviewed / Open Access
• [Presentation] Therapeutic effects of a novel RyR1 inhibitor on malignant hyperthermia model mice (2020)
  • Author(s): Murayama T, Yamazawa T, Kobayashi T, Kurebayashi N, Noguchi S, Nishino N, Mori S, Kagechika H, Lopez JR, Allen PD, Sakurai T
  • Organizer: Biophysical Society 64th Annual Meeting
• [Presentation] 新規RyR1抑制薬の悪性高熱症モデルマウスに対する効果 (2020)
  • Author(s): 村山 尚，山澤 徳志子，小林 琢也，呉林 なごみ，野口 悟，西野 一三，森 修一，影近 弘之，Jose R. Lopez，Paul D. Allen，櫻井 隆
  • Organizer: 第93回日本薬理学会年会
• [Presentation] 悪性高熱症モデルマウスを用いた新規RyR1阻害薬の治療効果の検討 (2019)
  • Author(s): 村山 尚、山澤 徳志子、小林 琢也、呉林 なごみ、野口 悟、井上 高良、井上 由紀子、森 修一、飯沼 大翔、間中 紀暁、湯浅 磨里、影近 弘之、西野 一三、櫻井 隆
  • Organizer: 第5回日本筋学会学術集会
• [Presentation] リアノジン受容体チャネル制御機構への構造・機能学的アプローチ (2019)
  • Author(s): 村山 尚
  • Organizer: 第92回日本生化学会大会
• [Presentation] Identification of novel inhibitors of Ca2+-induced Ca2+ release for RyR1-related muscle diseases (2019)
  • Author(s): Takashi Murayama
  • Organizer: 9th FAOPS Congress
  • Int'l Joint Research
• [Presentation] 小胞体内Ca2+濃度測定による新規RyR1阻害薬のハイスループットスクリーニング (2018)
  • Author(s): 村山 尚、呉林 なごみ、湯浅（石上） 磨里、森 修一、鈴木 志奈、秋間 龍之介、小川 治夫、鈴木 純二、金丸 和典、小山田 英人、木内 祐二、飯野 正光、影近 弘之、櫻井 隆
  • Organizer: 第4回日本筋学会
• [Presentation] 小胞体内Ca2+濃度測定によるリアノジン受容体阻害薬探索システムの開発 (2018)
  • Author(s): 村山 尚、呉林 なごみ、湯浅（石上） 磨里、森 修一、鈴木 志奈、秋間 龍之介、小川 治夫、鈴木 純二、金丸 和典、小山田 英人、木内 祐二、飯野 正光、影近 弘之、櫻井 隆
  • Organizer: 第139回日本薬理学会関東部会
• [Presentation] Molecular basis of Ca2+ binding to the ryanodine receptor for channel activation (2018)
  • Author(s): Murayama T, Ogawa H, Kurebayashi N, Ohno S, Horie M, Sakurai T
  • Organizer: 18th World Congress of Basic and Clinical Pharmacology
  • Int'l Joint Research
• [Presentation] Genotype-phenotype correlations of central core disease mutations in the C-terminal region of the RYR1 channel (2017)
  • Author(s): Takashi Murayama, Nagomi Kurebayashi, Haruo Ogawa, Toshiko Yamazawa, Takashi Sakurai
  • Organizer: Biophysical Society 61st Annual Meeting
• [Presentation] 1型リアノジン受容体（RYR1）チャネルC末端領域セントラルコア病変異の遺伝子型-表現型相関 (2017)
  • Author(s): 村山 尚、呉林なごみ、小川治夫、山澤徳志子、小山田英人、小口勝司、櫻井 隆
  • Organizer: 第90回日本薬理学会年会
• [Presentation] RyR1関連筋疾患の分子機構：診断と治療に向けて (2017)
  • Author(s): 村山 尚
  • Organizer: 第94回日本生理学会大会
• [Presentation] リアノジン受容体関連疾患の分子機構：診断と治療に向けて (2017)
  • Author(s): 村山 尚
  • Organizer: 第3回日本筋学会学術集会
• [Presentation] Molecular basis for Ca2+ and caffeine action on the RyR2 channel and implications in disease states (2017)
  • Author(s): Takashi Murayama, Haruo Ogawa, Nagomi Kurebayashi, Takashi Sakurai
  • Organizer: 20th International Symposium on Calcium Binding Proteins and Calcium Function in Health and Disease (CaBP20)
  • Int'l Joint Research
• [Presentation] Molecular mechanism of type 1 ryanodine receptor-related muscle diseases: toward diagnosis and therapy (2017)
  • Author(s): Takashi Murayama
  • Organizer: AuPS Scientific Meeting
  • Invited
• [Presentation] 1型リアノジン受容体C末端領域疾患変異体の機能解析 (2016)
  • Author(s): 村山 尚、呉林なごみ、山澤徳志子、小川治夫、小山田英人、小口勝司、櫻井 隆
  • Organizer: 第2回日本筋学会学術集会
  • Place of Presentation: 小平
  • Year and Date: 2016-08-05
• [Patent(Industrial Property Rights)] ２型リアノジン受容体活性抑制剤 (2020)
  • Inventor(s): 國廣なごみ、村山尚、影近弘之、森 修一、湯浅磨里、飯沼大翔
  • Industrial Property Rights Holder: 学校法人順天堂、国立大学法人東京医科歯科大学
  • Industrial Property Rights Type: 特許
  • Filing Date: 2020
• [Patent(Industrial Property Rights)] リアノジン受容体阻害薬 (2018)
  • Inventor(s): 村山 尚、國廣 なごみ、影近 弘之、森 修一、湯浅 磨里
  • Industrial Property Rights Holder: 村山 尚、國廣 なごみ、影近 弘之、森 修一、湯浅 磨里
  • Industrial Property Rights Type: 特許
  • Filing Date: 2018
  • Overseas
• [Patent(Industrial Property Rights)] リアノジン受容体阻害薬 (2017)
  • Inventor(s): 村山 尚、國廣 なごみ、影近 弘之、森 修一、湯浅 磨里
  • Industrial Property Rights Holder: 学校法人順天堂、国立大学法人東京医科歯科大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2017-205935
  • Filing Date: 2017
• [Patent(Industrial Property Rights)] Ｃａ2+誘発性Ｃａ2+遊離活性抑制剤のスクリーニング方法 (2016)
  • Inventor(s): 村山 尚、國廣 なごみ
  • Industrial Property Rights Holder: 順天堂大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2016-113147
  • Filing Date: 2016-06-07