title: Membrane potential-controlled inhibition of cytochrome c oxidase by zinc.
abstract: Like many voltage-sensitive ion pumps, cytochrome c oxidase is inhibited by zinc. Binding of zinc to the outside surface of Rhodobacter sphaeroides cytochrome c oxidase inhibits the enzyme with a K(I) of < or = 5 microm when the enzyme is reconstituted into phospholipid vesicles in the presence of a membrane potential. In the absence of a membrane potential and a pH gradient, millimolar concentrations of zinc are required to inhibit. This differential inhibition causes a dramatic increase in the respiratory control ratio from 6 to 40 for wild-type oxidase. The external zinc inhibition is removed by EDTA and is not competitive with cytochrome c binding but is competitive with protons. Only Cd(2+) of the many metals tested (Mg(2+), Mn(2+), Ca(2+), Ba(2+), Li(2+), Cs(2+), Hg(2+), Ni(2+), Co(2+), Cu(2+) Tb(3+), Tm(3+)) showed inhibitory effects similar to Zn(2+). Proton pumping is slower and less efficient with zinc. The results suggest that zinc inhibits proton movement through a proton exit path, which can allow proton back-leak at high membrane potentials. The physiological and mechanistic significance of proton movement in the exit pathway and its blockage by zinc is discussed in terms of regulation of the efficiency of energy transduction.
authors.affiliation: Department of Biochemistry, Michigan State University, East Lansing, Michigan 48824-1319, USA. millsden@pilot.msu.edu
authors.forename: Denise A
authors.initials: DA
authors.lastname: Mills
authors.forename: Bryan
authors.initials: B
authors.lastname: Schmidt
authors.forename: Carrie
authors.initials: C
authors.lastname: Hiser
authors.forename: Erica
authors.initials: E
authors.lastname: Westley
authors.forename: Shelagh
authors.initials: S
authors.lastname: Ferguson-Miller
mesh_terms: D003576:Electron Transport Complex IV / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D008564:Membrane Potentials; D008958:Models, Molecular; D011487:Protein Conformation; D012242:Rhodobacter sphaeroides / Q000201:enzymology; D015032:Zinc / Q000494:pharmacology*
chemical_list: D003576:Electron Transport Complex IV; D015032:Zinc