Fig. 10.

Demonstration of heteromeric CYP1A2•CYP2B4 complexes in reconstituted systems. Four distinct reconstituted systems were generated, two simple systems containing either CYP1A2 or CYP2B4, a combination of the separate vesicle systems (1A2 + 2B4), or a mixed reconstituted system containing equimolar concentrations of CYP1A2 and CYP2B4 in the same vesicles. These systems were first cross-linked with bis(sulfosuccinimidyl) suberate, then immunoprecipitated with CYP1A2 antibody, and finally immunoblotted with anti-CYP2B4 antibody. Higher molecular weight complexes (labeled as dimer, trimer, and tetramer) were observed only when both P450 enzymes were present in a common membrane. The “1A2 antibody” lane is a control generated without any reconstituted system. The molecular weight standard (MW std) has the masses of the bands indicated. Adapted from Reed et al., 2010.