Characterization of cytochrome c-556 from the purple phototrophic bacterium Rhodobacter capsulatus as a cytochrome-c peroxidase.
A cytochrome c-556 was purified from Rhodobacter capsulatus and the complete amino acid sequence was determined. It contains 328 amino acid residues and two typical heme-binding sites at cysteine residues 54 and 57 and at residues 200 and 203. It is homologous to the family of bacterial cytochrome c peroxidases (BCCP) with 69% identity to Paracoccus denitrificans BCCP and 60% identity to Pseudomonas aeruginosa BCCP for which there is a three-dimensional structure. There is lesser similarity to the mauG gene products from methylotrophic bacteria which are thought to be involved in biosynthesis of the quinone cofactor of methylamine dehydrogenase. Translated genes from Escherichia coli and Helicobacter pylori are also related to the bacterial cytochrome c peroxidases. The divergence of this family of proteins is reflected in the fact that the reported sixth heme ligands are not conserved, except in Pseudomonas, Rhodobacter and Paracoccus. This suggests that homologs of BCCP may fold differently and/or may not have the same enzymatic activity as the prototypic protein from Ps. aeruginosa. We found that the Rb. capsulatus BCCP is active with both Rb. capsulatus cytochrome c2 and with horse cytochrome c as substrates (Km values 60 microm and 6 microm, respectively). The turnover number was 40 s(-1) and the Km for peroxide was 33 microm. We have thus confirmed that the Rb. capsulatus protein is a cytochrome c peroxidase.
Hu
W
W
Department of Biochemistry, Physiology and Microbiology, University of Gent, Belgium.
De Smet
L
L
Van Driessche
G
G
Bartsch
R G
RG
Meyer
T E
TE
Cusanovich
M A
MA
Van Beeumen
J
J
D000595:Amino Acid Sequence; D003574:Cytochrome c Group / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D003578:Cytochrome-c Peroxidase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D016354:Rhodobacter capsulatus / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid
D003574:Cytochrome c Group; C020267:cytochrome c556; D003578:Cytochrome-c Peroxidase