Epitopes described in "Structure at 2.7 A resolution of the Paracoccus denitrificans two-subunit cytochrome c oxidase complexed with an antibody FV fragment."

Article Authors:C Ostermeier; A Harrenga; U Ermler; H Michel
Article Title:Structure at 2.7 A resolution of the Paracoccus denitrificans two-subunit cytochrome c oxidase complexed with an antibody FV fragment.
Reference Detail
Reference ID:1001550
Abstract:The aa3 type cytochrome c oxidase consisting of the core subunits I and II only was isolated from the soil bacterium Paracoccus denitrificans and crystallized as complex with a monoclonal antibody Fv fragment. Crystals could be grown in the presence of a number of different nonionic detergents. However, only undecyl-beta-D-maltoside and cyclohexyl-hexyl-beta-D-maltoside yielded well-ordered crystals suitable for high resolution x-ray crystallographic studies. The crystals belong to space group P212121 and diffract x-rays to at least 2.5 A (1 A = 0.1 nm) resolution using synchrotron radiation. The structure was determined to a resolution of 2.7 A using molecular replacement and refined to a crystallographic R-factor of 20.5% (Rfree = 25.9%). The refined model includes subunits I and II and the 2 chains of the Fv fragment, 2 heme A molecules, 3 copper atoms, and 1 Mg/Mn atom, a new metal (Ca) binding site, 52 tentatively identified water molecules, and 9 detergent molecules. Only four of the water molecules are located in the cytoplasmic half of cytochrome c oxidase. Most of them are near the interface of subunits I and II. Several waters form a hydrogen-bonded cluster, including the heme propionates and the Mg/Mn binding site. The Fv fragment binds to the periplasmic polar domain of subunit II and is critically involved in the formation of the crystal lattice. The crystallization procedure is well reproducible and will allow for the analysis of the structures of mechanistically interesting mutant cytochrome c oxidases.
Affiliations:Max-Planck-Institut für Biophysik, Abteilung für Molekulare Membranbiologie, Heinrich-Hoffmann-Strasse 7, D-60528 Frankfurt, Germany.
Reference Type:Literature
PubMed ID:9380672
Journal:Proc Natl Acad Sci U S A
Journal Volume:94
Article Pages:10547-53
Journal ISSN:0027-8424
Article Chemical List:Immunoglobulin Fragments;Metals;Protons;Electron Transport Complex IV
Article MeSH List:Binding Sites; Crystallography, X-Ray; Electron Transport Complex IV(chemistry; metabolism); Immunoglobulin Fragments(chemistry); Metals; Models, Chemical; Molecular Sequence Data; Paracoccus denitrificans(enzymology); Protein Conformation; Protons
Curation Last Updated:2015-06-05 00:27:08