Figure 1
Activation of E. coli cytochrome bd by CL. (A) Enzymatic function of cytochrome bd (PDB ID: 6RKO42, figure created with BioRender52). (B) The effect of CL (10 μM) on oxygen consumption activity by cytochrome bd (final conc. 2 nM) purified from E. coli was determined using a Clark-type electrode. The reaction was initiated by addition of ubiquinone-1 + DTT (arrow), the negative control contained ubiquinone-1 and DTT, but no cytochrome bd. Inset: Dependency of activation on the pre-incubation time at the indicated CL concentrations. The enzyme was incubated with CL prior to starting the reaction for either 3 min (black bars) or 60 min (white bars). (C) Impact of the inhibitor aurachin D (400 nM) on oxygen consumption by cytochrome bd in the presence or absence of 10 µM CL. (D) Effect of CL on the KM value of E. coli cytochrome bd. Curve fit was done with a simple Michaelis–Menten analysis; R2 values in the absence and presence of CL were 0.978 and 0.969, respectively. Average values were calculated from at least two biological replicates at 37 °C; error bars represent standard deviations.
