A comparative analysis of the active site properties of the resting states of cytochrome c peroxidase, metmyoglobin, and catalase

Abstract

AbstractQuantum chemical studies (INDO‐RHF‐SCF) have been made for the resting state active sites of three closely related heme proteins, cytochrome c peroxidase (CCP), metmyoglobin (MMB), and catalase (CAT). The relative energies of the germane sextet, quartet, and doublet spin‐states of each active site were calculated. Both CCP and MMB have similar heme units, consisting of an Fe(III)‐protoporphyrin‐IX with an imidazole and water as axial ligands. Our calculations show that the larger doming of the porphyrin, greater out‐of‐planarity of the iron, and the shorter iron‐water distance in MMB leads to a sextet ground state with a low‐lying quartet state. By contrast, the order of these two states is reversed in CCP, when a neutral imidazole is used as the endogenous axial ligand. An imidazolate ligand, on the other hand, which is an extreme representation of the H‐bonding believed to occur in CCP with a nearby aspartate residue, leads to a sextet ground state with a low‐lying quartet state. Assuming at least a partially anionic ligand in the intact protein, it follows that the quartet contribution to the ground state properties will be larger in CCP than in MMB. These predictions are consistent with the observed differences in the temperature‐dependent magnetic susceptibility for these two proteins. The present results suggest that the experimentally observed Mössbauer resonance spectra of CCP should be reinterpreted in terms of sextet and quartet state contributions to the electric field gradient. Calculations for catalase, which has a single phenolate ligand, result in a sextet ground state with a low‐lying quartet state consistent with available Mössbauer and magnetic susceptibility data. Our calculations of the Im− form of CCP show that it more closely resembles CAT. Thus, the effect of proton transfer in CCP can account at least in part for the similarities between CCP and CAT function. Minor differences in ground spin‐state and electronic properties calculated for CCP and MMB, however, cannot explain why MMB does not have significant peroxidase activity. The different functions of MMB and CCP must then be due in part to other known differences in their protein environment such as polar residues around the distal ligand binding pocket of CCP, which are absent in MMB, and could help its transformation to an active oxidizing state.