Characterization of the dihemic cytochrome C549 from the marine denitrifying bacterium Pseudomonas nautica 617

A dihemic ferricytochrome c549 (21 kDa) was purified and characterized from cells of the marine denitrifier Pseudomonas nautica strain 617. Several spectroscopic techniques, including UV-visible, NMR and EPR spectroscopies were applied to the characterization of this cytochrome. The visible and the 1H-NMR spectra show that both hemes have histidine-methionine as axial ligands. The dihemic cytochrome c549 has mid-point redox potentials of +230 mV and +250 mV, at pH 7.6 and its NH2-terminal sequence presents a high degree of similarity with those of cytochromes c4. The EPR studies allowed the determination of the orientation between the two axial ligands, indicating an axial ligand field for one of the hemes of cytochrome c549 and a rhombic symmetry for the other heme.