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. Author manuscript; available in PMC: 2007 Sep 24.
Published in final edited form as: Biochemistry. 2005 Feb 22;44(7):2632–2641. doi: 10.1021/bi0477900

Figure 4. Effects of ionic strength on PROD as a function of reductase concentration.

Figure 4

Pre-incubation of the binary systems containing 0.05 μM for both CYP2B4 and CYP1A2 in both the binary and ternary systems are described in Methods. The reductase concentration was varied as indicated. Groups represent the mean ± SEM for three determinations. The experimental data were fit to a model where CYP1A2 and CYP2B4 form a complex that influences reductase binding and are shown as a dotted line (Scheme 2). The dashed line shows the best fit based on model only allowing the formation of reductase-CYP1A2 and reductase-CYP2B4 complexes (Scheme 1). (a) Effect of mixed reconstitution of CYP1A2 and CYP2B4 on the reductase dependence of PROD at 50 mM HEPES. Concentrations of reductase were varied at the lower ionic strength (50 mM HEPES) buffer for CYP1A2, CYP2B4 and the mixed system containing both P450 enzymes. (b) Effect of mixed reconstitution of CYP1A2 and CYP2B4 on the reductase dependence of PROD (300 mM HEPES).