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测试去污剂用于大肠杆菌膜蛋白提取和脱脂效用的方案。

Protocol to test the utility of detergents for E. coli membrane protein extraction and delipidation.

作者信息

Wycisk Virginia, Urner Leonhard H

机构信息

TU Dortmund University, Department of Chemistry and Chemical Biology, Otto-Hahn-Str. 6, 44227 Dortmund, Germany.

TU Dortmund University, Department of Chemistry and Chemical Biology, Otto-Hahn-Str. 6, 44227 Dortmund, Germany.

出版信息

STAR Protoc. 2023 Mar 17;4(2):102146. doi: 10.1016/j.xpro.2023.102146.

Abstract

We present a protocol to evaluate the utility of detergents for purification and delipidation of E. coli membrane proteins. We determine the critical aggregation concentration of detergents. Furthermore, we compare the ability of detergents to extract membrane proteins and to maintain protein-lipid interactions during purification. The protocol describes steps for isolating and delipidating membrane proteins from E. coli membranes by extraction and affinity purification using detergents. The protocol does not enable an absolute quantification of purification outcomes. For complete details on the use and execution of this protocol, please refer to Urner et al..

摘要

我们提出了一种评估去污剂对大肠杆菌膜蛋白进行纯化和脱脂效用的方案。我们测定了去污剂的临界聚集浓度。此外,我们比较了去污剂在纯化过程中提取膜蛋白以及维持蛋白质-脂质相互作用的能力。该方案描述了使用去污剂通过提取和亲和纯化从大肠杆菌膜中分离和脱脂膜蛋白的步骤。该方案无法对纯化结果进行绝对定量。有关此方案的使用和执行的完整详细信息,请参考厄纳等人的研究。

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