SoxAX cytochromes, a new type of heme copper protein involved in bacterial energy generation from sulfur compounds.
SoxAX cytochromes are essential for the function of the only confirmed pathway for bacterial thiosulfate oxidation, the so-called "Sox pathway," in which they catalyze the initial formation of a S-S bond between thiosulfate and the SoxYZ carrier protein. Our work using the Starkeya novella diheme SoxAX protein reveals for the first time that in addition to two active site heme groups, SoxAX contains a mononuclear Cu(II) center with a distorted tetragonal geometry and three to four nitrogen ligands, one of which is a histidine. The Cu(II) center enhanced SoxAX activity in a newly developed, glutathione-based assay system that mimics the natural reaction of SoxAX with SoxYZ. EPR spectroscopy confirmed that the SoxAX Cu(II) center is reduced by glutathione. At pH 7 a K(m) (app) of 0.19+/-0.028 mm and a k(cat) (app) of 5.7+/-0.25s(-1) were determined for glutathione. We propose that SoxAX cytochromes are a new type of heme-copper proteins, with SoxAX-mediated S-S bond formation involving both the copper and heme centers.
Kappler
Ulrike
U
School of Molecular & Microbial Sciences, The University of Queensland, Brisbane, Queensland 4072, Australia. u.kappler@uq.edu.au
Bernhardt
Paul V
PV
Kilmartin
James
J
Riley
Mark J
MJ
Teschner
Julia
J
McKenzie
Kirsty J
KJ
Hanson
Graeme R
GR
D020561:Alphaproteobacteria / Q000201:enzymology / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D003067:Coenzymes / Q000378:metabolism; D003300:Copper / Q000378:metabolism*; D003574:Cytochrome c Group / Q000737:chemistry / Q000378:metabolism*; D004734:Energy Metabolism; D005978:Glutathione / Q000378:metabolism; D006418:Heme / Q000378:metabolism*; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013057:Spectrum Analysis; D013457:Sulfur Compounds / Q000378:metabolism*
D001426:Bacterial Proteins; D003067:Coenzymes; D003574:Cytochrome c Group; D011994:Recombinant Proteins; D013457:Sulfur Compounds; D006418:Heme; D003300:Copper; D010088:Oxidoreductases; D005978:Glutathione