Cytochrome c′ from Paracoccus denitrificans: spectroscopic studies consistent with a role for the protein in nitric oxide metabolism

Abstract

Cytochrome c′ was purified from the denitrifying bacterium Paracoccus denitrificans and the interaction of the protein with nitric oxide was examined spectroscopically. Two distinct types of haem-nitrosyl electronic absorption spectrum were observed, which were dependent upon [NO]. When cytochrome c′ was saturated with NO, α and β bands were centred at 562 nm and 530 nm, whereas with sub-saturating concentrations of NO the α and β bands were red-shifted to 578 nm and 542 nm respectively. Further spectroscopic analysis showed that purified cytochrome c′, added to suspensions of P. denitrificans, is able to complex with the NO which is formed as a freely diffusible intermediate of denitrification. In the presence of added NO − 3 or NO − 2, 40–60% of Fe(II)-cytochrome c′ forms a 6-coordinate haem-nitrosyl complex. In the absence of nitrogen oxyanions or NO whole denitrifying cells are able to remove the NO from a Fe(II)-cytochrome c′-NO complex. These findings support the hypothesis that the physiological function of this enigmatic cytochrome involves the reversible binding of nitric oxide.