Programmed axon death executor SARM1 is a multi-functional NAD(P)ase with prominent base exchange activity, all regulated by physiological levels of NMN, NAD, NADP and other metabolites

SARM1 is an NAD glycohydrolase and TLR adapter with an essential, prodegenerative role in programmed axon death (Wallerian degeneration). It has low basal NADase activity that becomes strongly activated by NAD precursor NMN. Very high levels of NAD oppose this activation, competing for the same allosteric site on SARM1’s regulatory ARM domain. Injury or diseases that deplete axons of NMNAT2, an essential enzyme converting NMN to NAD, cause SARM1 activation. The resulting NAD degradation by SARM1, combined with loss of NAD synthesis by NMNAT2, causes rapid depletion of axonal NAD. This NAD loss is widely assumed to mediate axon death and is consequently a key focus for therapeutic strategies for axonopathies. However, like other NAD(P) glycohydrolases, SARM1 has additional enzyme activities whose contributions, consequences and regulation need to be fully understood. Here, we compare the multiple actions and regulation of recombinant human SARM1 with those of two other NAD(P) glycohydrolases, human CD38 and Aplysia californica ADP ribosyl cyclase. We find that SARM1 has the highest transglycosidation (base exchange) activity of these enzymes at neutral pH and with some bases this dominates NAD(P) hydrolysis and cyclisation. Moreover, like its NADase and NADPase reactions, SARM1-mediated base exchange at neutral pH is activated by increases in the NMN:NAD ratio, which we show for the first time can act in the presence of physiological levels of both metabolites. We establish that SARM1 base exchange is the most likely physiological source of calcium mobilizing agent NaADP, and potentially of other NAD(P) analogues, which could contribute to axon and cell death. We also identify regulatory effects of free pyridine bases, of NADP and of nicotinic acid riboside (NaR) on SARM1 that represent further therapeutic opportunities. These data will help to pinpoint which of the multiple functions of SARM1 is responsible for axon degeneration and how it can be optimally targeted to block axon degeneration in disease. ### Competing Interest Statement Funding for academic research from AstraZeneca and M.P.C. is a consultant for Nura Bio. * #### ABBREVIATIONS ARM : armadillo/HEAT motif NAD(P) : nicotinamide adenine dinucleotide (phosphate) oxidized forms; this is used throughout to mean NAD+ or NADP+; NaAD(P) : nicotinic acid adenine dinucleotide (phosphate) oxidized forms; this is used throughout to mean NaAD+ or NaADP+; NaR : nicotinic acid riboside NR : nicotinamide riboside NAD(P)ase : NAD(P) glycohydrolase (EC 3.2.2.6) NMNAT2 : nicotinamide mononucleotide adenylyltransferase (EC 2.7.7.1) isoform 2 NMN : nicotinamide mononucleotide ROS : reactive oxygen species SAM : sterile-α motif SARM1 : sterile alpha and TIR motif-containing protein 1 TIR : Toll/interleukin-1 receptor; vacor or pyrinuron, 1-(4-nitrophenyl)-3-(pyridin-3-ylmethyl)urea VAD : vacor adenine dinucleotide VMN : vacor mononucleotide VR : vacor riboside.