Extended Data Fig. 5: NADH turnover by PYROXD1.
(a) Single-turnover kinetics of NADH oxidation by human PYROXD1 variants under aerobic conditions. Data points represent the mean ± SEM of three independent replicates. Solid lines represent a single-exponential fit. (b) Multiple-turnover kinetics of NADH oxidation by human PYROXD1 variants under aerobic conditions. Data points represent the mean ± SEM of three independent replicates. Solid lines represent a linear fit. (c) Michealis-Menten analysis of NADH oxidation by human WT and W239A PYROXD1 under aerobic conditions. Data points represent the mean ± SEM of three independent replicates. Solid lines represent a Michaelis-Menten fit. (d) Co-precipitation of PYROXD1 mutants by immobilized StrepII-GFP-RTCB in the presence of NADH or NADPH. Bound proteins were analyzed by SDS-PAGE and Coomassie blue staining. This experiment was repeated three times with similar results.
