细菌的主动膜转运和受体蛋白。

Active membrane transport and receptor proteins from bacteria.

作者信息

Saidijam M, Bettaney K E, Szakonyi G, Psakis G, Shibayama K, Suzuki S, Clough J L, Blessie V, Abu-Bakr A, Baumberg S, Meuller J, Hoyle C K, Palmer S L, Butaye P, Walravens K, Patching S G, O'reilly J, Rutherford N G, Bill R M, Roper D I, Phillips-Jones M K, Henderson P J F

机构信息

Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, U.K.

出版信息

Biochem Soc Trans. 2005 Aug;33(Pt 4):867-72. doi: 10.1042/BST0330867.

DOI:10.1042/BST0330867

PMID:16042616

Abstract

A general strategy for the expression of bacterial membrane transport and receptor genes in Escherichia coli is described. Expression is amplified so that the encoded proteins comprise 5-35% of E. coli inner membrane protein. Depending upon their topology, proteins are produced with RGSH6 or a Strep tag at the C-terminus. These enable purification in mg quantities for crystallization and NMR studies. Examples of one nutrient uptake and one multidrug extrusion protein from Helicobacter pylori are described. This strategy is successful for membrane proteins from H. pylori, E. coli, Enterococcus faecalis, Bacillus subtilis, Staphylococcus aureus, Microbacterium liquefaciens, Brucella abortus, Brucella melitensis, Campylobacter jejuni, Neisseria meningitides, Streptomyces coelicolor and Rhodobacter sphaeroides.

摘要

本文描述了一种在大肠杆菌中表达细菌膜转运和受体基因的通用策略。表达量被放大，使得编码的蛋白质占大肠杆菌内膜蛋白的5%-35%。根据其拓扑结构，蛋白质在C端带有RGSH6或链霉亲和素标签。这使得能够以毫克量进行纯化，用于结晶和核磁共振研究。文中描述了来自幽门螺杆菌的一种营养物质摄取蛋白和一种多药外排蛋白的实例。该策略对于来自幽门螺杆菌、大肠杆菌、粪肠球菌、枯草芽孢杆菌、金黄色葡萄球菌、液化微杆菌、流产布鲁氏菌、羊种布鲁氏菌、空肠弯曲菌、脑膜炎奈瑟菌、天蓝色链霉菌和球形红杆菌的膜蛋白均取得成功。