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  A D-Pathway Mutation Decouples the Paracoccus denitrificans Cytochrome c Oxidase by Altering the Side-Chain Orientation of a Distant Conserved Glutamate

Dürr, K. L., Koepke, J., Hellwig, P., Müller, H., Angerer, H., Peng, G., et al. (2008). A D-Pathway Mutation Decouples the Paracoccus denitrificans Cytochrome c Oxidase by Altering the Side-Chain Orientation of a Distant Conserved Glutamate. Journal of Molecular Biology (London), 384(4), 865-877. doi:10.1016/j.jmb.2008.09.074.

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 Creators:
Dürr, Katharina L.1, Author           
Koepke, Jürgen2, Author           
Hellwig, Petra1, 3, Author
Müller, Hannelore2, Author           
Angerer, Heike2, Author           
Peng, Guohong2, Author           
Olkhova, Elena2, Author           
Richter, Oliver-Matthias H.1, Author
Ludwig, Bernd1, Author
Michel, Hartmut2, 4, Author           
Affiliations:
1Department of Biochemistry, Molecular Genetics Group, Johann Wolfgang Goethe University, 60438 Frankfurt/Main, Germany, ou_persistent22              
2Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society, ou_2068290              
3Institute of Biophysics, Johann Wolfgang Goethe University, 60438 Frankfurt/Main, Germany, ou_persistent22              
4Cluster of Excellence Frankfurt Macromolecular Complexes, Johann Wolfgang Goethe University, 60438 Frankfurt/Main, Germany, ou_persistent22              

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Free keywords: Proton pumping; proton gating; water chain; electron transfer; FTIR spectroscopy
 Abstract: Asparagine 131, located near the cytoplasmic entrance of the D-pathway in subunit I of the Paracoccus denitrificans aa3 cytochrome c oxidase, is a residue crucial for proton pumping. When replaced by an aspartate, the mutant enzyme is completely decoupled: while retaining full cytochrome c oxidation activity, it does not pump protons. The same phenotype is observed for two other substitutions at this position (N131E and N131C), whereas a conservative replacement by glutamine affects both activities of the enzyme. The N131D variant oxidase was crystallized and its structure was solved to 2.32-Å resolution, revealing no significant overall change in the protein structure when compared with the wild type (WT), except for an alternative orientation of the E278 side chain in addition to its WT conformation. Moreover, remarkable differences in the crystallographically resolved chain of water molecules in the D-pathway are found for the variant: four water molecules that are observed in the water chain between N131 and E278 in the WT structure are not visible in the variant, indicating a higher mobility of these water molecules. Electrochemically induced Fourier transform infrared difference spectra of decoupled mutants confirm that the protonation state of E278 is unaltered by these mutations but indicate a distinct perturbation in the hydrogen-bonding environment of this residue. Furthermore, they suggest that the carboxylate side chain of the N131D mutant is deprotonated. These findings are discussed in terms of their mechanistic implications for proton routing through the D-pathway of cytochrome c oxidase.

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Language(s): eng - English
 Dates: 2008-08-282008-06-272008-09-172008-10-092008
 Publication Status: Published in print
 Pages: 13
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1016/j.jmb.2008.09.074
 Degree: -

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Title: Journal of Molecular Biology (London)
  Other : J Mol Biol
Source Genre: Journal
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Publ. Info: London : Academic Press
Pages: - Volume / Issue: 384 (4) Sequence Number: - Start / End Page: 865 - 877 Identifier: ISSN: 0022-2836
CoNE: https://pure.mpg.de/cone/journals/resource/954922646042