Biochemical and spectroscopic characterization of two new cytochromes isolated from Desulfuromonas acetoxidans.
The multimeric cytochromes described to date in sulfate- and sulfur-reducing bacteria are associated with diverse respiratory modes involving the use of elemental sulfur or oxidized sulfur compounds as terminal acceptors. They exhibit no structural similarity with the other cytochrome c classes and are characterized by a bis-histidinyl axial iron coordination and low redox potentials. We have purified two new cytochromes c with markedly different molecular masses (10 000 and 50 000) from the bacterium Desulfuromonas acetoxidans, which uses anaerobic sulfur respiration as its sole energy source. The characterization by electrochemistry and optical and EPR spectroscopies revealed the cytochrome c (Mr = 10 000) to be the first monohemic cytochrome c exhibiting a bis-histidinyl axial coordination and a low redox potential (-220 mV). The cytochrome c (Mr = 50 000) contains four hemes of low potential (-200, -210, -370, and -380 mV) with the same axial coordination. The N-terminal amino acid sequences were compared with that of the trihemic cytochrome c7, previously described in D. acetoxidans and which is related to tetrahemic cytochrome c3 from sulfate reducing bacteria. Some homology was found between cytochrome c (Mr = 10 000) and cytochrome c7. Both D. acetoxidans cytochromes c are located in the periplasmic space and their biochemical and spectroscopic properties indicate that they belong to the class III cytochromes.
Bruschi
M
M
Laboratoire de Bioénergétique et Ingénierie des Protéines, IFR 1, C.N.R.S., Marseille, France. bruschi@ibsm.cnrs-mrs.fr
Woudstra
M
M
Guigliarelli
B
B
Asso
M
M
Lojou
E
E
Petillot
Y
Y
Abergel
C
C
D000596:Amino Acids / Q000032:analysis; D003574:Cytochrome c Group / Q000737:chemistry* / Q000302:isolation & purification*; D016969:Desulfovibrio vulgaris / Q000201:enzymology; D004563:Electrochemistry; D004578:Electron Spin Resonance Spectroscopy; D006089:Gram-Negative Anaerobic Bacteria / Q000201:enzymology; D006418:Heme / Q000737:chemistry; D007501:Iron / Q000737:chemistry; D008969:Molecular Sequence Data; D016999:Sulfur-Reducing Bacteria / Q000201:enzymology*; D017186:Titrimetry
D000596:Amino Acids; D003574:Cytochrome c Group; D006418:Heme; D007501:Iron