Proton diffusion along biological membranes is vitally important for cellular energetics. Here we extended previous time-resolved fluorescence measurements to study the time and temperature dependence of surface proton transport. We determined the Gibbs activation energy barrier ΔG ‡ r that opposes proton surface-to-bulk release from Arrhenius plots of (i) protons’ surface diffusion constant and (ii) the rate coefficient for proton surface-to-bulk release. The large size of ΔG ‡ r disproves that quasi-equilibrium exists in our experiments between protons in the near-membrane layers and in the aqueous bulk. Instead, non-equilibrium kinetics describes the proton travel between the site of its photo-release and its arrival at a distant membrane patch at different temperatures. ΔG ‡ r contains only a minor enthalpic contribution that roughly corresponds to the breakage of a single hydrogen bond. Thus, our experiments reveal an entropic trap that ensures channeling of highly mobile protons along the membrane interface in the absence of potent acceptors.

Respiratory complex I catalyzes electron transfer from NADH to ubiquinone through a chain of seven iron-sulfur (Fe-S) clusters in the hydrophilic domain. Four protons are transported across the membrane contributing to the electrochemical proton gradient required for ATP synthesis. The electron transfer properties of the Fe-S clusters in Thermus thermophilus complex I are investigated here using the Density Functional Theory þ Poisson-Boltzmann (DFTþPB) approach. The reduction potentials of the Fe-S clusters are calculated to give insight into the reduction potential energy profile of the main redox chain.

Respiratory complex I catalyzes electron transfer from NADH to ubiquinone through a chain of seven iron-sulfur (Fe-S) clusters in the hydrophilic domain. Four protons are transported across the membrane contributing to the electrochemical proton gradient required for ATP synthesis. The electron transfer properties of the Fe-S clusters in Thermus thermophilus complex I are investigated here using the Density Functional Theory þ Poisson-Boltzmann (DFTþPB) approach. The reduction potentials of the Fe-S clusters are calculated to give insight into the reduction potential energy profile of the main redox chain.