Free energy profiles for H+ conduction in the D-pathway of Cytochrome c Oxidase: a study of the wild type and N98D mutant enzymes.
The molecular mechanism for proton conduction in the D-pathway of Cytochrome c Oxidase (CcO) is investigated through the free energy profile, i.e., potential of mean force (PMF) calculations of both the native enzyme and the N98D mutant. The multistate empirical valence bond (MS-EVB) model was applied to simulate the interaction of an excess proton with the channel environment. In the study of the wild type enzyme, the PMF reveals the previously proposed proton trap inside the channel; it also shows a high free energy barrier against the passage of proton at the entry of the channel, where two conserved asparagines (ASN80/98) may be essential for the gating of proton uptake. We also present data from an investigation of the N98D mutant, which has been previously shown to completely eliminate proton pumping but significantly enhance the oxidase activity in Rhodobacter sphaeroides. These results suggest that mutating Asn98 to negatively charged aspartate will create an unfavorable energy barrier sufficiently high to prevent the overall proton uptake through the D-pathway, whereas with a protonated aspartic acid the proton conduction was found to be accelerated. Plausible explanations for the origin of the uncoupling of proton pumping from the oxidase activity will be discussed.
Xu
Jiancong
J
Department of Chemistry, University of Utah, Salt Lake City, 84112-0850, USA.
Voth
Gregory A
GA
D000818:Animals; D002417:Cattle; D003576:Electron Transport Complex IV / Q000235:genetics / Q000378:metabolism*; D006859:Hydrogen / Q000378:metabolism*; D009206:Myocardium / Q000201:enzymology; D017354:Point Mutation; D013816:Thermodynamics
D006859:Hydrogen; D003576:Electron Transport Complex IV