A conformation-specific nanobody targeting the nicotinamide mononucleotide-activated state of SARM1

Yun Nan Hou; Yang Cai; Wan Hua Li; Wei Ming He; Zhi Ying Zhao; Wen Jie Zhu; Qiang Wang; Xinyi Mai; Jun Liu; Hon Cheung Lee; Goran Stjepanovic; Hongmin Zhang; Yong Juan Zhao

doi:10.1038/s41467-022-35581-y

A conformation-specific nanobody targeting the nicotinamide mononucleotide-activated state of SARM1

Nat Commun. 2022 Dec 22;13(1):7898. doi: 10.1038/s41467-022-35581-y.

Authors

Yun Nan Hou^#¹, Yang Cai^#², Wan Hua Li^#^{1

3}, Wei Ming He¹, Zhi Ying Zhao¹, Wen Jie Zhu¹, Qiang Wang¹, Xinyi Mai⁴, Jun Liu¹, Hon Cheung Lee¹, Goran Stjepanovic⁵, Hongmin Zhang⁶, Yong Juan Zhao^{7

8}

Affiliations

¹ State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, 518055, China.
² Department of Biology, School of Life Sciences, Southern University of Science and Technology, Shenzhen, 518055, China.
³ Ciechanover Institute of Precision and Regenerative Medicine, School of Medicine, The Chinese University of Hong Kong, Shenzhen, 518172, China.
⁴ Kobilka Institute of Innovative Drug Discovery, School of Medicine, The Chinese University of Hong Kong, Shenzhen, 518172, China.
⁵ Kobilka Institute of Innovative Drug Discovery, School of Medicine, The Chinese University of Hong Kong, Shenzhen, 518172, China. goranstjepanovic@cuhk.edu.cn.
⁶ Department of Biology, School of Life Sciences, Southern University of Science and Technology, Shenzhen, 518055, China. zhanghm@sustech.edu.cn.
⁷ State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, 518055, China. zhaoyongjuan@cuhk.edu.cn.
⁸ Ciechanover Institute of Precision and Regenerative Medicine, School of Medicine, The Chinese University of Hong Kong, Shenzhen, 518172, China. zhaoyongjuan@cuhk.edu.cn.

^# Contributed equally.

Abstract

Sterile alpha (SAM) and Toll/interleukin-1 receptor (TIR) motif containing 1 (SARM1) is an autoinhibitory NAD-consuming enzyme that is activated by the accumulation of nicotinamide mononucleotide (NMN) during axonal injury. Its activation mechanism is not fully understood. Here, we generate a nanobody, Nb-C6, that specifically recognizes NMN-activated SARM1. Nb-C6 stains only the activated SARM1 in cells stimulated with CZ-48, a permeant mimetic of NMN, and partially activates SARM1 in vitro and in cells. Cryo-EM of NMN/SARM1/Nb-C6 complex shows an octameric structure with ARM domains bending significantly inward and swinging out together with TIR domains. Nb-C6 binds to SAM domain of the activated SARM1 and stabilized its ARM domain. Mass spectrometry analyses indicate that the activated SARM1 in solution is highly dynamic and that the neighboring TIRs form transient dimers via the surface close to one BB loop. We show that Nb-C6 is a valuable tool for studies of SARM1 activation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Armadillo Domain Proteins / genetics
Armadillo Domain Proteins / metabolism
Axons* / metabolism
Nicotinamide Mononucleotide* / metabolism
Protein Domains

Substances

Nicotinamide Mononucleotide
Armadillo Domain Proteins