Permeant fluorescent probes visualize the activation of SARM1 and uncover an anti-neurodegenerative drug candidate
- The Chinese University of Hong Kong (Shenzhen), China
- Hong Kong Baptist University, China
- Southern University of Science and Technology, China
- Peking University Shenzhen Graduate School, China
Abstract
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 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.
Data availability
Diffraction data have been deposited in PDB under the accession code 7DJT. All data generated or analysed during this study are included in the manuscript and supporting files.
Article and author information
Author details
Funding
Ministry of Science and Technology of the People's Republic of China (2019YFA090600)
- Hongmin Zhang
National Science Foundation of China (31871401)
- Yong Juan Zhao
Hong Kong Baptist University (RC-SGT2/18-19/SCI/005)
- Chi-Sing Lee
Hong Kong Baptist University (RC-ICRS-18-19-01A)
- Chi-Sing Lee
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: This study was carried out in strict accordance with animal use protocol approved by Peking University Shenzhen Graduate School Animal Care and Use Committee (#AP0015001). All animals (C57BL6/J), purchased from Guangdong Medical Laboratory Animal Center (China), were handled in accordance with the guidelines of the Committee on the Ethic of Animal Experiments. All surgery was performed after euthanasia and efforts were made to minimize suffering.
Copyright
© 2021, Li et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Permeant fluorescent probes visualize the activation of SARM1 and uncover an anti-neurodegenerative drug candidate
eLife 10:e67381.
https://doi.org/10.7554/eLife.67381
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