Directed evolution of a bacterial efflux pump:: Adaptation of the E-coli TolC exit duct to the Pseudomonas MexAB translocase

被引：50

作者：

Bokma, Evert ^{[1
]}

Koronakis, Eva ^{[1
]}

Lobedanz, Sune ^{[1
]}

Hughes, Colin ^{[1
]}

Koronakis, Vassilis ^{[1
]}

机构：

[1] Univ Cambridge, Dept Pathol, Cambridge CB2 1QP, England

来源：

FEBS LETTERS | 2006年 / 580卷 / 22期

基金：

英国惠康基金;

关键词：

membrane protein; multidrug resistance; efflux pump;

D O I：

10.1016/j.febslet.2006.09.005

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Bacterial multidrug efflux pumps operate by periplasmic recruitment and opening of TolC family outer membrane exit ducts by cognate inner membrane translocases. Directed evolution of active hybrid pumps was achieved by challenging a library of mutated, shuffled TolC variants to adapt to the noncognate Pseudomonas MexAB translocase, and confer resistance to the efflux substrate novobiocin. Amino acid substitutions in MexAB-adapted TolC variants that endowed high resistance were recreated independently, and revealed that MexAB-adaptation was conferred only by substitutions located in the lower a-helical barrel of TolC, specifically the periplasmic equatorial domain and entrance coiled coils. These changes converge to the native MexAB partner OprM, and indicate an interface key to the function and diversity of efflux pumps. (c) 2006 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

引用

页码：5339 / 5343

页数：5

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