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. Author manuscript; available in PMC: 2018 Dec 7.
Published in final edited form as: Neuron. 2017 Mar 22;93(6):1334–1343.e5. doi: 10.1016/j.neuron.2017.02.022

Figure 3: Characterization of the SARM1-TIR NAD+ cleavage reaction.

Figure 3:

(A-E) HPLC chromatograms showing NAD+ cleavage products of human and drosophila SARM1-TIR. Retention time: Nam t~2.40 min; cADPR at t~0.85 min; ADPR at t~1.10 min. (F-G) Quantification of metabolites generated by human (F) and drosophila (G) SARM1-TIR as displayed in A-E (normalized to 0 min NAD+). H) Kinetic parameters for human SARM1-TIR cleavage reaction. Vmax, Km, kcat were determined by fitting the data to the Michaelis-Menten equation and are presented as mean ± SEM for three independent biological samples. I) Nam dose response inhibition of human SARM1-TIR enzymatic activity. Data was generated from three independent reaction experiments using purified protein from three independent bacteria clones. Data are presented as mean ± SEM; Error bars: SEM; *P<0.05; **P<0.01; ***P<0.001 unpaired two tailed Student’s t-test. See also Figure S3.