A Water Dimer Shift Activates a Proton Pumping Pathway in the PR → F Transition of ba3 Cytochrome c Oxidase

Abstract

Broken-symmetry density functional calculations have been performed on the [Fe_a3⁴⁺,Cu_B²⁺] state of the dinuclear center (DNC) for the P_R → F part of the catalytic cycle of ba₃ cytochrome c oxidase (CcO) from Thermus thermophilus (Tt), using the OLYP-D3-BJ functional. The calculations show that the movement of the H₂O molecules in the DNC affects the pK_a values of the residue side chains of Tyr237 and His376⁺, which are crucial for proton transfer/pumping in ba₃ CcO from Tt. The calculated lowest energy structure of the DNC in the [Fe_a3⁴⁺,Cu_B²⁺] state (state F) is of the form Fe_a3⁴⁺═O^2-···Cu_B²⁺, in which the H₂O ligand that resulted from protonation of the OH^- ligand in the P_R state is dissociated from the Cu_B²⁺ site. The calculated Fe_a3⁴⁺═O^2- distance in F (1.68 Å) is 0.03 Å longer than that in P_R (1.65 Å), which can explain the different Fe_a3⁴⁺═O^2- stretching modes in P (804 cm^-1) and F (785 cm^-1) identified by resonance Raman experiments. In this F state, the Cu_B²⁺···O^2- (ferryl-oxygen) distance is only around 2.4 Å. Hence, the subsequent O_H state [Fe_a3³⁺-OH^--Cu_B²⁺] with a μ-hydroxo bridge can be easily formed, as shown by our calculations.