Structure analysis of the membrane protein TatCd from the Tat system of B. subtilis by circular dichroism
Structure analysis of the membrane protein TatCd from the Tat system of B. subtilis by circular dichroism
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DOI：
10.1016/j.bbamem.2009.07.003
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发表时间：
2009-10-01
影响因子：
3.4
通讯作者：
Ulrich, Anne S.
中科院分区：
文献类型：
--
作者：
Nolandt, Olga V.;Walther, Torsten H.;Ulrich, Anne S.
双精氨酸易位（达特）系统可以运输完全折叠的蛋白质，包括它们的辅因子，穿过细菌和类囊体膜。用于输出磷酸二酯酶（PhoD）的枯草芽孢杆菌的达特系统仅由两种膜蛋白TatA（d）和TatC（d）组成。较大的组分TatCd具有28 kDa的分子量和几个跨膜片段。这种蛋白质已在大肠杆菌中表达，并纯化到足够的量，用于圆二色性（CD）和NMR光谱的结构分析。将TatC（d）在洗涤剂胶束和脂质双层中重构，用于溶液和宏观定向样品中的CID分析，以检查蛋白质的稳定性。已经建立了合适的协议和模型膜系统，通过这些协议和模型膜系统，TatC（d）保持螺旋度接近理论预测的水平，并且可以验证其跨膜排列。(C)2009爱思唯尔有限公司版权所有。
The twin arginine translocation (Tat) system can transport fully folded proteins, including their cofactors, across bacterial and thylakoid membranes. The Tat system of Bacillus subtilis that serves to export the phosphodiesterase (PhoD) consists of only two membrane proteins, TatA(d) and TatC(d). The larger component TatCd has a molecular weight of 28 kDa and several membrane-spanning segments. This protein has been expressed in Escherichia coli and purified in sufficient amounts for structure analysis by circular dichroism (CD) and NMR spectroscopy. TatC(d) was reconstituted in detergent micelles and in lipid bilayers for CID analysis in solution and in macroscopically oriented samples, to examine the stability of the protein. Suitable protocols and model membrane systems have been established, by which TatC(d) maintains the level of helicity close to theoretically predicted, and its transmembrane alignment could been verified. (C) 2009 Elsevier B.V. All rights reserved.