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来自嗜盐栖热菌的渗透调节型四氢嘧啶转运蛋白TeaABC的底物结合蛋白TeaA:重组TeaA的纯化与特性分析

The substrate-binding protein TeaA of the osmoregulated ectoine transporter TeaABC from Halomonas elongata: purification and characterization of recombinant TeaA.

作者信息

Tetsch Larissa, Kunte Hans Jörg

机构信息

Institut für Mikrobiologie und Biotechnologie, Rheinische-Friedrich-Wilhelms-Universität, Meckenheimer Allee 168, D-53115, Bonn, Germany.

出版信息

FEMS Microbiol Lett. 2002 Jun 4;211(2):213-8. doi: 10.1111/j.1574-6968.2002.tb11227.x.

Abstract

TeaABC is a novel, osmoregulated secondary transport system for compatible solutes found in the halophilic proteobacterium Halomonas elongata DSM 2581(T). Sequence comparisons have confirmed that TeaABC belongs to the family of TRAP transporters and as such it consists of two putative transmembrane proteins (TeaB, TeaC) and a putative periplasmic substrate-binding protein (TeaA). TeaABC is the only osmoregulated transporter for ectoines found in H. elongata. By overexpressing TeaA-(6)His tag in Escherichia coli we demonstrated that TeaA is processed as predicted and exported to the periplasm. Furthermore, it was proven that TeaA is indeed a periplasmic ectoine-binding protein. The functionality of the purified protein as an ectoine-binding protein was tested employing a modified binding assay.

摘要

TeaABC是一种新型的、渗透调节的次级转运系统,用于转运嗜盐变形杆菌长盐单胞菌DSM 2581(T)中发现的相容性溶质。序列比较证实,TeaABC属于TRAP转运蛋白家族,因此它由两个假定的跨膜蛋白(TeaB、TeaC)和一个假定的周质底物结合蛋白(TeaA)组成。TeaABC是长盐单胞菌中唯一发现的用于转运ectoine的渗透调节转运蛋白。通过在大肠杆菌中过表达带有(6)His标签的TeaA,我们证明TeaA按预测进行加工并输出到周质。此外,已证明TeaA确实是一种周质ectoine结合蛋白。采用改良的结合试验测试了纯化蛋白作为ectoine结合蛋白的功能。

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