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          Institute: MPI für Biophysik     Collection: Abt. Molekulare Membranbiologie     Display Documents

ID: 680367.0, MPI für Biophysik / Abt. Molekulare Membranbiologie
A decade of crystallization drops: Crystallization of the cbb3 cytochrome c oxidase from Pseudomonas stutzeri
Authors:Buschmann, Sabine; Richers, Sebastian; Ermler, Ulrich; Michel, Hartmut
Date of Publication (YYYY-MM-DD):2014-04
Title of Journal:Protein Science
Journal Abbrev.:Protein Sci.
Issue / Number:4
Start Page:411
End Page:422
Review Status:Peer-review
Audience:Experts Only
Abstract / Description:The cbb3 cytochrome c oxidases are distant members of the superfamily of heme copper oxidases. These terminal oxidases couple O2 reduction with proton transport across the plasma membrane and, as a part of the respiratory chain, contribute to the generation of an electrochemical proton gradient. Compared with other structurally characterized members of the heme copper oxidases, the recently determined cbb3 oxidase structure at 3.2 Å resolution revealed significant differences in the electron supply system, the proton conducting pathways and the coupling of O2 reduction to proton translocation. In this paper, we present a detailed report on the key steps for structure determination. Improvement of the protein quality was achieved by optimization of the number of lipids attached to the protein as well as the separation of two cbb3 oxidase isoenzymes. The exchange of n-dodecyl-β-D-maltoside for a precisely defined mixture of two α-maltosides and decanoylsucrose as well as the choice of the crystallization method had a most profound impact on crystal quality. This report highlights problems frequently encountered in membrane protein crystallization and offers meaningful approaches to improve crystal quality.
Free Keywords:cbb3 oxidase; membrane protein; crystallization; lipid content; controlled delipidation; detergent
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für Biophysik/Abteilung Molekulare Membranbiologie
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