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TitleA conformation-specific nanobody targeting the nicotinamide mononucleotide-activated state of SARM1.
Journal, issue, pagesNat Commun, Vol. 13, Issue 1, Page 7898, Year 2022
Publish dateDec 22, 2022
AuthorsYun 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 /
PubMed AbstractSterile 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 ...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.
External linksNat Commun / PubMed:36550129 / PubMed Central
MethodsEM (single particle)
Resolution2.7 - 3.78 Å
Structure data

34165

8gni

EMDB-34165, PDB-8gni:
Human SARM1 bounded with NMN and Nanobody-C6, Conformation 1
Method: EM (single particle) / Resolution: 3.74 Å

34166

8gnj

EMDB-34166, PDB-8gnj:
Human SARM1 bounded with NMN and Nanobody-C6, Conformation 2
Method: EM (single particle) / Resolution: 3.78 Å

34198

8gq5

EMDB-34198, PDB-8gq5:
Human SARM1 bounded with NMN and Nanobody-C6, double-layer structure
Method: EM (single particle) / Resolution: 2.7 Å

Chemicals

ChemComp-NMN:
BETA-NICOTINAMIDE RIBOSE MONOPHOSPHATE / Nicotinamide mononucleotide

Source
  • homo sapiens (human)
  • vicugna pacos (alpaca)
KeywordsHYDROLASE / NAD(+)Hydrolase / NMN / Nanobody

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