Resonance Raman study of deoxy and ligated (O2 and CO) mesoheme IX-reconstituted myoglobin, hemoglobin and its alpha and beta subunits.

In this work, we corrected the resonance Raman (RR) results presented earlier for deoxy mesoheme IX-reconstituted hemoglobin (mesoHb) alpha and beta subunits implied that mesohemes in these subunits undergo substantial structural changes upon formation of a hemoglobin tetramer (Biochemistry 29 (1990) 5087). We show that these data were probably due to the improper handling of the deoxy mesoheme subunit preparation. Additionally, we discuss the RR spectra of deoxy, oxy, and CO species of mesoheme IX-reconstituted myoglobin (mesoMb) and alpha and beta deoxy meso hemoglobin subunits, including their analogues with deuterium-substituted mesoheme IX in all methyl groups (d(12)). Based on the obtained data, we propose a complete RR band assignment for all of the investigated molecules. The most pronounced changes are observed for the gamma(7) mode (out-of-plane movement of methane carbon atoms) associated with the interaction of the ethyl groups with the globin. We also show that in mesoheme IX-reconstituted proteins, the O(2) molecule binds stronger than in the case of native species. This is manifested by the up-shift of nu(Fe-O(2)).