2. Academic Validation
  3. Crystal structures of Ryanodine Receptor reveal dantrolene and azumolene interactions guiding inhibitor development

Crystal structures of Ryanodine Receptor reveal dantrolene and azumolene interactions guiding inhibitor development

  • Nat Commun. 2025 Nov 18;16(1):10110. doi: 10.1038/s41467-025-65096-1.
Hadiatullah Hadiatullah # 1,  Lianyun Lin # 1,  Zhiyan Wang # 2,  Rajamanikandan Sundarraj 1,  Qing Wang 1,  Xinru Lai 1,  Nagomi Kurebayashi 3,  Takuya Kobayashi 3,  Toshiko Yamazawa 4,  Yu Seby Chen 5,  Wenlan Wang 1,  Hongxia Zhao 1,  Yiqing Yin 6,  Takashi Murayama 3,  Filip Van Petegem 5,  Zhiguang Yuchi 7 8 9 10
Affiliations

Affiliations

  • 1 State Key Laboratory of Synthetic Biology; Frontiers Science Center for Synthetic Biology; Tianjin Key Laboratory for Modern Drug Delivery & High-Efficiency; School of Pharmaceutical Science and Technology, Faculty of Medicine, Tianjin University, Tianjin, China.
  • 2 Department of Molecular Pharmacology, Tianjin Medical University Cancer Institute & Hospital; National Clinical Research Center for Cancer; Key Laboratory of Cancer Prevention and Therapy; Tianjin's Clinical Research Center for Cancer, Tianjin, China.
  • 3 Department of Pharmacology, Juntendo University School of Medicine, Tokyo, Japan.
  • 4 Core Research Facilities, The Jikei University School of Medicine, Tokyo, Japan.
  • 5 Department of Biochemistry and Molecular Biology, Life Sciences Institute, University of British Columbia, Vancouver, BC, Canada.
  • 6 Department of Anesthesiology, Tianjin Medical University Cancer Institute & Hospital; National Clinical Research Center for Cancer; Key Laboratory of Cancer Prevention and Therapy; Tianjin's Clinical Research Center for Cancer, Tianjin, China.
  • 7 State Key Laboratory of Synthetic Biology; Frontiers Science Center for Synthetic Biology; Tianjin Key Laboratory for Modern Drug Delivery & High-Efficiency; School of Pharmaceutical Science and Technology, Faculty of Medicine, Tianjin University, Tianjin, China. [email protected].
  • 8 Department of Molecular Pharmacology, Tianjin Medical University Cancer Institute & Hospital; National Clinical Research Center for Cancer; Key Laboratory of Cancer Prevention and Therapy; Tianjin's Clinical Research Center for Cancer, Tianjin, China. [email protected].
  • 9 Haihe Laboratory of Sustainable Chemical Transformations, Tianjin, China. [email protected].
  • 10 Guangdong Laboratory for Lingnan Modern Agriculture (Shenzhen Branch), Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen, Guangdong, China. [email protected].
  • # Contributed equally.
PMID: 41253812 DOI: 10.1038/s41467-025-65096-1
Abstract

The ryanodine receptor (RyR) is a critical drug target, yet dantrolene (DAN) remains the only FDA-approved inhibitor, limited by hepatotoxicity and unsuitable for chronic use. To guide improved inhibitor development, we determine high-resolution crystal structures of the RyR Repeat12 (R12) domain bound to DAN, its analog azumolene (AZU), and adenine nucleotides (AMP-PCP or ADP). DAN/AZU and nucleotides bind cooperatively to a pseudosymmetric cleft, with key interactions involving Trp880 and Trp994. Binding induces a clamshell-like closure of the R12 domain. Isothermal titration calorimetry (ITC) reveals higher affinity in the presence of nucleotides and lower affinity for RyR2 due to nearby substitutions. Structural comparison with cryo-EM data suggests that DAN/AZU binding allosterically influences RyR gating and functional regulation. Structure-based screening identifies a potent compound targeting the same site but with a distinct binding mode. Our findings highlight the power of domain-focused crystallography in guiding RyR inhibitor discovery and overcoming cryo-EM resolution limitations.

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