The reaction of Escherichia coli cytochrome bo with H2O2: evidence for the formation of an oxyferryl species by two distinct routes.
We have re-examined the reaction of fast oxidised cytochrome bo with H202 in a stopped-flow spectrophotometer. Monitoring the reaction at 582 nm allows us to observe the formation and decay of a spectroscopically distinct intermediate which accumulates transiently prior to the formation of an oxyferryl species previously characterised in this laboratory (Watmough, N.J., Cheesman, M.R., Greenwood, C. and Thomson, A.J. (1994) Biochem. J. 300, 469-475 [1]). The reaction shows three distinct phases of which the fast and intermediate phases are bimolecular and show a marked pH dependence. Initially these results appeared incompatible with the report that only one equivalent of H202 is required to generate the oxyferryl species (Moody, A.J. and Rich, P.R. (1994) Eur. J. Biochem. 226, 731-737 [21]. However, these data can be reconciled by a branched reaction mechanism whose contributions differ according to the peroxide concentration used.
Brittain
T
T
Centre for Metalloprotein Spectroscopy and Biology, School of Biological Sciences, University of East Anglia, Norwich, UK.
Little
R H
RH
Greenwood
C
C
Watmough
N J
NJ
D003573:Cytochrome b Group; D003580:Cytochromes / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins; D005290:Ferric Compounds / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics
D003573:Cytochrome b Group; D003580:Cytochromes; D029968:Escherichia coli Proteins; D005290:Ferric Compounds; C017168:cytochrome bo, E coli; D006861:Hydrogen Peroxide