科研之友
微信
新浪微博
Facebook
分享链接
摘要
The cbb(3) cytochrome c oxidases are distant members of the superfamily of heme copper oxidases. These terminal oxidases couple O-2 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 cbb(3) oxidase structure at 3.2 angstrom resolution revealed significant differences in the electron supply system, the proton conducting pathways and the coupling of O-2 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 cbb(3) oxidase isoenzymes. The exchange of n-dodecyl--d-maltoside for a precisely defined mixture of two -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.
- 出版日期2014-4
