A Conformation-specific Nanobody Targeting the Nicotinamide Mononucleotide-activated State of SARM1

Overview

Journal Nat Commun

Specialty Biology

Date 2022 Dec 22

PMID 36550129

Authors

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

Affiliations

Soon will be listed here.

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.

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