Fig. 6

Single-wavelength stopped-flow experiments. a Stopped-flow single wavelength absorbance (443 nm) after mixing 20 μM of GSTACMO with 200 μM of NADPH or 4(R)-[4-²H]NADPH. Absorbance traces (black lines) were collected over 1 s (a) and fitted to standard single exponential equation (grey dotted line), which gave values of k_red1 of 28.4 ± 0.5 s⁻¹ (NADPH) and 7.5 ± 0.2 s⁻¹ (4(R)-[4-²H]NADPH). b Dependence of k_red1 on the concentration of NADPH of wild type ACMO (solid circles) and the H325K variant (open circles). The data were fitted to Eq. 6 and the results are shown in Table 2