Permeant fluorescent probes visualize the activation of SARM1 and uncover an anti-neurodegenerative drug candidate

eLife

Ke Huang¹  Qian Wen Wang²  Yong Juan Zhao²  Hon Cheung Lee³  Yang Cai³  Chi-Sing Lee⁴  Zhi Ying Zhao⁴  Yang Du⁵  Wan Hua Li⁵  Hongmin Zhang⁵  Sujing Wang⁵  Yun Nan Hou⁵  Xu Jie Xie⁵  Wen Jie Zhu⁵  Sheng Cao⁵ 

[1] Ciechanover Institute of Precision and Regenerative Medicine, School of Life and Health Sciences, The Chinese University of Hong Kong, Shenzhen, China;Department of Biology, Southern University of Science and Technology, Shenzhen, China;Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong, China;Kobilka Institute of Innovative Drug Discovery, School of Life and Health Sciences, The Chinese University of Hong Kong, Shenzhen, China;State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, China;

关键词: SARM1;    fluorescent probes;    base-exchange;    allosteric inhibitors;    CryoEM structure;    covalent inhibitors;   

DOI  :  10.7554/eLife.67381

来源: DOAJ

【 摘 要 】

SARM1 regulates axonal degeneration through its NAD-metabolizing activity and is a drug target for neurodegenerative disorders. We designed and synthesized fluorescent conjugates of styryl derivative with pyridine to serve as substrates of SARM1, which exhibited large red shifts after conversion. With the conjugates, SARM1 activation was visualized in live cells following elevation of endogenous NMN or treatment with a cell-permeant NMN-analog. In neurons, imaging documented mouse SARM1 activation preceded vincristine-induced axonal degeneration by hours. Library screening identified a derivative of nisoldipine (NSDP) as a covalent inhibitor of SARM1 that reacted with the cysteines, especially Cys311 in its ARM domain and blocked its NMN-activation, protecting axons from degeneration. The Cryo-EM structure showed that SARM1 was locked into an inactive conformation by the inhibitor, uncovering a potential neuroprotective mechanism of dihydropyridines.

【 授权许可】

Unknown