Structural similarities and differences of the heme pockets of various P450 isoforms as revealed by resonance Raman spectroscopy.

Cytochromes P450 CYP102 A1, 1A2, and 3A4, all belonging to the class II type of P450 enzymes, were studied by resonance Raman spectroscopy. Spectra were measured for the oxidized substrate-free, oxidized substrate-bound, and reduced forms of each of these P450s. The analysis of the resonance Raman spectra indicates that the individual isoforms differ with respect to orientation and conformations of the heme side chains, whereas the overall porphyrin geometry is essentially the same. In the oxidized state, the vinyl groups exhibit both a coplanar and an out-of-plane orientation with respect to the heme, albeit with different relative propensities in the various isoforms. In the reduced state, both vinyl groups are forced into a coplanar orientation. In addition to the differences in behavior of the vinyl groups, the redox-linked spectral changes also include the bending mode of the propionate side chains. The spectral differences associated with the porphyrin substituents are likely to reflect subtle conformational differences in the heme pocket of various P450 isoforms which may constitute the structural basis for the known variability of their functions.