Effect of chemical modification of lysine amino groups on redox and protonmotive activity of bovine heart cytochrome c oxidase reconstituted in phospholipid membranes.
A study is presented of the effect of chemical modification of lysine amino groups on the redox and protonmotive activity of bovine heart cytochrome c oxidase. Treatment of soluble oxidase with succinic acid anhydride resulted in succinylation of lysines in all the subunits of the enzyme. The consequent change of surface charges from positive to negative resulted in inversion of the orientation of the reconstituted enzyme from right-side-out to inside-out. Reconstitution of the oxidase in phospholipid vesicles prevented succinylation of subunits III and Vb and depressed that of other subunits with the exception of subunits II and IV which were predominantly labeled in a concentration-dependent manner by succinic acid anhydride. This modification of lysines produced a decoupling effect on redox-linked proton ejection, which was associated with a decrease of the respiratory control exerted by the delta pH component of PMF. The decoupling effect was directly shown to be exerted at the level of the pH-dependent rate-limiting step in intramolecular electron flow located on the oxygen side of heme a.
Steverding
D
D
Institute of Medical Biochemistry and Chemistry, University of Bari, Italy.
Kadenbach
B
B
Capitanio
N
N
Papa
S
S
D000818:Animals; D002417:Cattle; D003576:Electron Transport Complex IV / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008081:Liposomes; D008239:Lysine; D046911:Macromolecular Substances; D008929:Mitochondria, Heart / Q000201:enzymology*; D009550:Nigericin / Q000494:pharmacology; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D010743:Phospholipids; D011485:Protein Binding; D011522:Protons; D013387:Succinic Anhydrides / Q000378:metabolism / Q000494:pharmacology; D014634:Valinomycin / Q000494:pharmacology
D008081:Liposomes; D046911:Macromolecular Substances; D010743:Phospholipids; D011522:Protons; D013387:Succinic Anhydrides; D014634:Valinomycin; C031801:succinic anhydride; D003576:Electron Transport Complex IV; D008239:Lysine; D009550:Nigericin