Zinc ions inhibit oxidation of cytochrome c oxidase by oxygen.
Cytochrome c oxidase is a membrane-bound enzyme that catalyses the reduction of O2 to H2O and uses part of the energy released in this reaction to pump protons across the membrane. We have investigated the effect of addition of Zn2+ on the kinetics of two reaction steps in cytochrome c oxidase that are associated with proton pumping; the peroxy to oxo-ferryl (P(r)-->F) and the oxo-ferryl to oxidised (F-->O) transitions. The Zn2+ binding resulted in a decrease of the F-->O rate from 820 s(-1) (no Zn2+) to a saturating value of approximately 360 s(-1) with an apparent K(D) of approximately 2.6 microM. The P(r)-->F rate (approximately 10[(4) s(-1)] before addition of Zn2+) decreased more slowly with increasing Zn2+ concentration and a K(D) of approximately 120 microM was observed. The effects on both kinetic phases were fully reversible upon addition of EDTA. Since both the P(r)-->F and F-->O transitions are associated with proton uptake through the D-pathway, a Zn2+-binding site is likely to be located at the entry point of this pathway, where several carboxylates and histidine residues are found that may co-ordinate Zn2+.
Aagaard
A
A
Department of Biochemistry and Biophysics, The Arrhenius Laboratories for Natural Sciences, Stockholm University, SE-106 91 Stockholm, Sweden.
Brzezinski
P
P
D001665:Binding Sites; D002384:Catalysis / Q000187:drug effects; D002413:Cations, Divalent / Q000378:metabolism / Q000494:pharmacology; D003576:Electron Transport Complex IV / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D006639:Histidine / Q000378:metabolism; D007700:Kinetics; D016877:Oxidants / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010100:Oxygen / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011487:Protein Conformation; D011522:Protons; D012242:Rhodobacter sphaeroides / Q000201:enzymology*; D015032:Zinc / Q000378:metabolism / Q000494:pharmacology*
D002413:Cations, Divalent; D016877:Oxidants; D011522:Protons; D006639:Histidine; D003576:Electron Transport Complex IV; D015032:Zinc; D010100:Oxygen