Structural and functional basis of the selectivity filter as a gate in human TRPM2 channel
Xiafei Yu,
Yuan Xie,
Xiaokang Zhang,
Cheng Ma,
Likun Liu,
Wenxuan Zhen,
Lingyi Xu,
Jianmin Zhang,
Yan Liang,
Lixia Zhao,
Xiuxia Gao,
Peilin Yu,
Jianhong Luo,
Lin-Hua Jiang,
Yan Nie,
Fan Yang,
Jiangtao Guo,
Wei Yang
Affiliations
Xiafei Yu
Departments of Biophysics and Neurosurgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, P. R. China
Yuan Xie
Departments of Biophysics and Pathology of Sir Run 14, Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310058, P. R. China
Xiaokang Zhang
Center of Cryo-Electron Microscopy, Zhejiang University School of Medicine, Hangzhou 310058, P. R. China; Interdisciplinary Center for Brain Information, The Brain Cognition and Brain Disease Institute, Faculty of Life and Health Sciences, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, Guangdong 518055, China; Shenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research Institutions, Shenzhen, Guangdong 518055, China
Cheng Ma
Co-facility center, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, P. R. China
Likun Liu
Departments of Biophysics and Neurosurgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, P. R. China
Wenxuan Zhen
Department of Biophysics and Kidney Disease Center, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, P. R. China
Lingyi Xu
Departments of Biophysics and Pathology of Sir Run 14, Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310058, P. R. China
Jianmin Zhang
Departments of Biophysics and Neurosurgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, P. R. China
Yan Liang
Departments of Toxicology and Medical Oncology of Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, P. R. China
Lixia Zhao
Shanghai Institute for Advanced Immunochemical Studies, ShanghaiTech University, Shanghai 201210, P. R. China
Xiuxia Gao
Shanghai Institute for Advanced Immunochemical Studies, ShanghaiTech University, Shanghai 201210, P. R. China
Peilin Yu
Departments of Toxicology and Medical Oncology of Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, P. R. China
Jianhong Luo
Department of Neurobiology, Key Laboratory of Medical Neurobiology of the Ministry of Health of China, Zhejiang Province Key Laboratory of Neurobiology, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, P. R. China
Lin-Hua Jiang
School of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UK; Sino-UK Joint Laboratory of Brain Function and Injury, Department of Physiology and Neurobiology, Xinxiang Medical University, Xinxiang, P. R. China
Yan Nie
Department of Neurobiology, Key Laboratory of Medical Neurobiology of the Ministry of Health of China, Zhejiang Province Key Laboratory of Neurobiology, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, P. R. China
Fan Yang
Department of Biophysics and Kidney Disease Center, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, P. R. China; Corresponding author
Jiangtao Guo
Departments of Biophysics and Pathology of Sir Run 14, Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310058, P. R. China; Corresponding author
Wei Yang
Departments of Biophysics and Neurosurgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, P. R. China; Corresponding author
Summary: Transient receptor potential melastatin 2 (TRPM2), a Ca2+-permeable cation channel, is gated by intracellular adenosine diphosphate ribose (ADPR), Ca2+, warm temperature, and oxidative stress. It is critically involved in physiological and pathological processes ranging from inflammation to stroke to neurodegeneration. At present, the channel’s gating and ion permeation mechanisms, such as the location and identity of the selectivity filter, remain ambiguous. Here, we report the cryo-electron microscopy (cryo-EM) structure of human TRPM2 in nanodisc in the ligand-free state. Cryo-EM map-guided computational modeling and patch-clamp recording further identify a quadruple-residue motif as the ion selectivity filter, which adopts a restrictive conformation in the closed state and acts as a gate, profoundly contrasting with its widely open conformation in the Nematostella vectensis TRPM2. Our study reveals the gating of human TRPM2 by the filter and demonstrates the feasibility of using cryo-EM in conjunction with computational modeling and functional studies to garner structural information for intrinsically dynamic but functionally important domains.
