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. 2021 May 18;296:100794. doi: 10.1016/j.jbc.2021.100794

Figure 3.

Figure 3

Effect of adrenodoxin on CYP11B2-mediated catalysis. Michaelis–Menten kinetics for CYP11B2-mediated metabolism of 11-deoxycorticosterone in the presence of 1-fold, 10-fold, and 40-fold excess of adrenodoxin (A) and for the adrenodoxin–CYP11B2 fusion enzyme with 40-fold excess of adrenodoxin (C). Rates were determined for corticosterone formation from 11-deoxycorticosterone. For experiments with 1-fold adrenodoxin, a CYP11B2 concentration of 0.2 μM was used. For experiments with 10-fold and 40-fold adrenodoxin, a reduced CYP11B2 concentration of 0.05 μM was used to prevent substrate depletion. For experiments with the adrenodoxin-CYP11B2 fusion protein, 0.4 μM protein concentration was used. Product profiles did not change under these conditions. Note the differences in the two y-axes. Data are shown in technical triplicates and was fit to the Michaelis–Menten equation to derive the kinetic parameters Km and kcat (B).