Water chain formation and possible proton pumping routes in Rhodobacter sphaeroides cytochrome c oxidase: a molecular dynamics comparison of the wild type and R481K mutant.
Cytochrome c oxidase (CcO) converts the energy from redox and oxygen chemistry to support proton translocation and create a transmembrane DeltamuH(+) used for ATP production. Molecular dynamics (MD) simulations were carried out to probe for the formation water chains capable of participating in proton translocation. Attention was focused on the region between and above the a and a(3) hemes where well-defined water chains have not been identified in crystallographic studies. An arginine (R481) (Rhodobacter sphaeroides numbering), positioned between the D-propionates of the hemes, had been mutated in vivo to lysine and showed to have altered activity consistent with an altered proton conductance [Qian, J., Mills, D. A., Geren, L., Wang, K. F., Hoganson, C. W., Schmidt, B., Hiser, C., Babcock, G. T., Durham, B., Millett, F., and Ferguson-Miller, S. (2004) Role of the conserved arginine pair in proton and electron transfer in cytochrome c oxidase, Biochemistry 43, 5748-5756; also see the accompanying paper by Mills et al.]. This mutant was created in silico, and the MD results for the mutant and wild type were compared to explore the effects on the formation of hydrogen-bonded water chains by this mutation. The simulations reveal the presence of hydrogen-bonded water chains that lead from E286 through the region above the hemes to the Mg(2+), and from E286 to the heme a(3) D-propionate and the binuclear center. The R481K mutant does not form as many, or as extensive, water chains as wild-type CcO, due to a new conformation of residues in a large loop between helices III and IV in subunit I, indicating a reduction in the level of water chain formation in the mutant. This loop appears to play a role in controlling the formation of hydrogen-bonded water chains above the hemes. The results suggest a possible gating mechanism for proton movement that includes key residues W172 and Y175 on the loop and F282 on helix VI.
Seibold
Steve A
SA
Department of Chemistry, Michigan State University, East Lansing, Michigan 48824, USA.
Mills
Denise A
DA
Ferguson-Miller
Shelagh
S
Cukier
Robert I
RI
D001120:Arginine / Q000235:genetics*; D003198:Computer Simulation; D003576:Electron Transport Complex IV / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D018698:Glutamic Acid / Q000737:chemistry; D006418:Heme / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D006860:Hydrogen Bonding; D008239:Lysine / Q000235:genetics*; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D011422:Propionates / Q000737:chemistry; D017433:Protein Structure, Secondary; D017494:Proton Pumps / Q000737:chemistry* / Q000378:metabolism; D012242:Rhodobacter sphaeroides / Q000201:enzymology* / Q000235:genetics; D013816:Thermodynamics; D014867:Water / Q000737:chemistry* / Q000378:metabolism
D011422:Propionates; D017494:Proton Pumps; D014867:Water; C027728:heme a; D018698:Glutamic Acid; D006418:Heme; D001120:Arginine; D003576:Electron Transport Complex IV; C029658:propionic acid; D008239:Lysine