Fig. 1: NUDT15 diphosphatase activity on nucleotide analogs.

a Twelve nucleotide analogs were tested for their reactivity with NUDT15, with TGTP (triphosphate nucleotide metabolite of thiopurine) as the reference. Corresponding prodrug names are shown below each metabolite in parenthesis. b Wild-type (WT) NUDT15 protein (200 ng) was incubated with various nucleotide analogs for 10 min followed by measuring released pyrophosphate (PPi) to evaluate the diphosphatase activity. Asterisks denote the value was below the detection limit. Data represent the mean of three replicates; error bars, s.d. c WT NUDT15 or p. R139C variant protein was subjected to diphosphatase activity measurement with acyclovir triphosphate (ACV-TP) or ganciclovir triphosphate (GCV-TP) as the substrate. NUDT15 p.R139C resulted in complete loss of enzymatic activity on both substrates. Data represent the mean of duplicates and experiments were repeated three times. d WT NUDT15 protein (200 ng) was incubated with 50 μM of either ACV-TP or GCV-TP for 3 min, and reaction samples were analyzed by the UPLC-MS/MS system. Representative data from three biological replicates are shown. ACV-TP and GCV-TP were converted to the monophosphates, ACV-MP and GCV-MP, respectively. Source data for b, c, and d are provided as a Source Data file.