Cytochrome ba3 from Natronobacterium pharaonis--an archaeal four-subunit cytochrome-c-type oxidase.

Cytochrome ba3, a terminal oxidase was isolated from the haloalkaliphilic archaeon Natronobacterium pharaonis. NH2-terminal sequence information of two subunits with apparent molecular masses of 40 and 36 kDa was used to generate a DNA probe by polymerase chain reaction. Cloning and sequencing of two overlapping genomic fragments revealed four genes forming a transcriptional unit. The policystronic messenger RNA of this cbaDBAC gene locus was identified by RNA analysis. The genes cbaC and cbaD code for small hydrophobic peptides with 81 and 54 amino acids. The genes cbaB and cbaA code for cytochrome oxidase subunit II (calculated molecular mass = 18.6 kDa) and I (calculated molecular mass = 63.8 kDa) respectively. Five potential CuA ligands for subunit II and six His residues for subunit I located in conserved positions indicate cytochrome ba3 to be a c-type oxidase. Sequence comparison and phylogenetic analysis place the natronobacterial enzyme together with the archaeal quinol oxidase SoxABCD from Sulfolobus acidocaldarius and the eubacterial ba3-type oxidase from Thermus thermophilus into a distinct evolutionary group. All three members are missing residues which are functionally important for vectorial proton translocation. The four-subunit enzyme complex was also identified on the protein level using chromatographic buffers containing ethylene glycol for purification.