多组分药物外排转运体的生化重组与表征
Biochemical Reconstitution and Characterization of Multicomponent Drug Efflux Transporters.
作者信息
Picard Martin, Tikhonova Elena B, Broutin Isabelle, Lu Shuo, Verchère Alice, Zgurskaya Helen I
机构信息
Laboratoire de Biologie Physico-chimique des ProtÕines Membranaires, UMR7099. IBPC, UniversitÕ Paris Diderot, 13, Rue Pierre et Marie Curie, 75005, Paris, France.
Department of Cell Biology and Biochemistry, Texas Tech University Health Science Center, 3601 4th Street, Lubbock, TX, 79430, USA.
出版信息
Methods Mol Biol. 2018;1700:113-145. doi: 10.1007/978-1-4939-7454-2_8.
Efflux pumps are the major determinants in bacterial multidrug resistance. In Gram-negative bacteria, efflux transporters are organized as macromolecular tripartite machineries that span the two-membrane cell envelope of the bacterium. Biochemical data on purified proteins are essential to draw a mechanistic picture of this highly dynamical, multicomponent, efflux system. We describe protocols for the reconstitution and the in vitro study of transporters belonging to RND and ABC superfamilies: the AcrAB-TolC and MacAB-TolC efflux systems from Escherichia coli and the MexAB-OprM efflux pump from Pseudomonas aeruginosa.
外排泵是细菌多药耐药性的主要决定因素。在革兰氏阴性菌中，外排转运蛋白组装成跨越细菌双膜细胞包膜的大分子三方机制。关于纯化蛋白的生化数据对于描绘这个高度动态、多组分的外排系统的机制图至关重要。我们描述了用于重组和体外研究属于RND和ABC超家族的转运蛋白的方案：来自大肠杆菌的AcrAB-TolC和MacAB-TolC外排系统以及来自铜绿假单胞菌的MexAB-OprM外排泵。
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