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- W2002122819 abstract "The 4-subunit crystal structure (PDB:1M56) of Rhodobacter sphaeroides cytochrome c oxidase (RsCcO) does not resolve the last six residues of the C-terminus of subunit I. In the higher resolution 2-subunit structure (PDB: 2GSM), ten more residues in the C-terminus are missing, suggesting flexibility of this region, which is also implied by deuterium exchange and computational analysis. To determine the functional significance of the C-terminus, three mutants of RsCcO were created, CIΔ16, CIΔ6 and E552A. The 16-residue deletion mutant, CIΔ16 (C-terminus = T550), showed suicide inactivation and lower steady state activity compared to wild type RsCcO. In the purified, reconstituted CIΔ16, proton pumping is greatly impaired, though its respiratory control is good. SDS-PAGE indicates that subunit III is partially lost. The shorter deletion mutant, CIΔ6 (C-terminus = W560), had normal activity and no suicide inactivation, suggesting that loss of activity in CIΔ16 could be due to the importance of some residue(s) in the 551-560 region. A conserved glutamate, E552, was a likely candidate, and in fact the E552A mutant had normal activity but diminished proton-pumping. Crystal structures of CIΔ16 and CIΔ6 were obtained at 2.1 Å and 2.5 Å respectively. The CIΔ16 structure shows a rearrangement of the new C-terminus (T550) to interact directly with H26, but the ordered waters above D132 in the D-pathway were unaltered. The CIΔ6 structure appeared WT. A role for E552 in proton pumping is implied, and of the 551 to 560 region in stabilizing the association of subunit I and III. (NIH GM26916)." @default.
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- W2002122819 date "2012-01-01" @default.
- W2002122819 modified "2023-09-27" @default.
- W2002122819 title "Crystallographic and Functional Evidence for Involvement of the Flexible C-Terminus of Subunit I of Rhodobacter Sphaeroides Cytochrome C Oxidase in Proton Pumping" @default.
- W2002122819 doi "https://doi.org/10.1016/j.bpj.2011.11.3108" @default.
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