Structural analysis of myeloperoxidase by resonance Raman spectroscopy.

Soret excitation of canine myeloperoxidase (MPO) gives rise to a complex resonance Raman (RR) spectrum characterized by multiple bands in the core size and oxidation state marker regions. Relative intensities of the bands obtained by 406- and 454-nm laser excitation were nearly identical and were temperature independent from 77 to 273 K. Spectra of dithionite-reduced and cyanide-coordinated derivatives are also reported. In the native and dithionite-reduced enzyme, there are no detectable bands between 1620 and 1700 cm-1, indicating that the hemes do not contain formyl substituents in conjugation with the macrocyclic ring. Excitation of the visible absorption band at 568 nm gave rise to only very weakly resonance-enhanced spectra. The RR spectra are interpreted within the context of other physical measurements to indicate that MPO contains two equivalent or nearly equivalent chlorin prosthetic groups. Possible mechanistic consequences of these structural features are discussed.