Flash-induced membrane potential generation by cytochrome c oxidase.
Flash-induced single-electron reduction of cytochrome c oxidase. Compound F (oxoferryl state) by RuII(2,2'-bipyridyl)3(2+) [Nilsson (1992) Proc. Natl. Acad. Sci. USA 89, 6497-6501] gives rise to three phases of membrane potential generation in proteoliposomes with tau values and contributions of ca. 45 microsecond (20%), 1 ms (20%) and 5 ms (60%). The rapid phase is not sensitive to the binuclear centre ligands, such as cyanide or peroxide, and is assigned to vectorial electron transfer from CuA to heme a. The two slow phases kinetically match reoxidation of heme a, require added H2O2 or methyl peroxide for full development, and are completely inhibited by cyanide; evidently, they are associated with the reduction of Compound F to the Ox state by heme a. The charge transfer steps associated with the F to Ox conversion are likely to comprise (i) electrogenic uptake of a 'chemical' proton from the N phase required for protonation of the reduced oxygen atom and (ii) electrogenic H+ pumping across the membrane linked to the F to Ox transition. Assuming heme a 'electrical location' in the middle of the dielectric barrier, the ratio of the rapid to slow electrogenic phase amplitudes indicates that the F to Ox transition is linked to transmembrane translocation of 1.5 charges (protons) in addition to an electrogenic uptake of one 'chemical' proton required to form Fe(3+)-OH- from Fe4+ = O2-. The shortfall in the number of pumped protons and the biphasic kinetics of the millisecond part of the electric response matching biphasic reoxidation of heme a may indicate the presence of 2 forms of Compound F, reduction of only one of which being linked to full proton pumping.
Zaslavsky
D
D
A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russian Federation.
Kaulen
A D
AD
Smirnova
I A
IA
Vygodina
T
T
Konstantinov
A A
AA
D000818:Animals; D002374:Catalase / Q000494:pharmacology; D002417:Cattle; D004563:Electrochemistry / Q000379:methods; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000378:metabolism*; D008027:Light; D008081:Liposomes; D008564:Membrane Potentials; D008929:Mitochondria, Heart / Q000201:enzymology; D008962:Models, Theoretical; D010713:Phosphatidylcholines; D010743:Phospholipids; D011190:Potassium Cyanide / Q000494:pharmacology; D011510:Proteolipids
D008081:Liposomes; D010713:Phosphatidylcholines; D010743:Phospholipids; D011510:Proteolipids; C020485:proteoliposomes; C024990:asolectin; D002374:Catalase; D003576:Electron Transport Complex IV; D011190:Potassium Cyanide