Deduction of consensus binding sequences on proteins that bind IIAGlc of the phosphoenolpyruvate:sugar phosphotransferase system by cysteine scanning mutagenesis of Escherichia coli lactose permease.
Deduction of consensus binding sequences on proteins that bind IIAGlc of the phosphoenolpyruvate:sugar phosphotransferase system by cysteine scanning mutagenesis of Escherichia coli lactose permease.
复制标题
通过大肠杆菌乳糖通透酶的半胱氨酸扫描诱变推导结合磷酸烯醇丙酮酸：糖磷酸转移酶系统的 IIAGlc 的蛋白质上的共有结合序列。
DOI：
10.1073/pnas.96.7.3525
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发表时间：
1999
影响因子：
11.1
通讯作者：
Peterkofsky,A
中科院分区：
文献类型：
--
作者：
Sondej,M;Sun,J;Seok,YJ;Kaback,HR;Peterkofsky,A
在大肠杆菌中，磷酸烯醇式丙酮酸：糖磷酸转移酶系统在蛋白IIAGlc的介导下，对其它糖转运系统的活性起调节作用。通过使用直接结合试验，对乳糖通透酶的胞质环中的单Cys置换突变体的集合评估其结合IIAGlc的能力。环IV/V或VI/VII中的选择性Cys置换导致结合活性的丧失。对诱变结果的分析以及与IIAGlc相互作用的蛋白质家族的多重序列比对为在那些结合IIAGlc所必需的伴侣蛋白中开发两个共有序列区域提供了基础。在底物依赖性构象变化的调控框架中解释了对两个相互作用区域的要求，所述底物依赖性构象变化使这两个区域进入结合IIAGlc的最佳方向。
Mediated by the protein IIAGlc, the phosphoenolpyruvate:sugar phosphotransferase system plays a role in the regulation of activity of other sugar transport systems inEscherichia coli. By using a direct binding assay, a collection of single-Cys replacement mutants in cytoplasmic loops of lactose permease were evaluated for their capacity to bind IIAGlc. Selected Cys replacements in loops IV/V or VI/VII result in loss of binding activity. Analysis of the mutagenesis results together with multiple sequence alignments of a family of proteins that interacts with IIAGlcprovides the basis for developing two regions of consensus sequence in those partner proteins necessary for binding to IIAGlc. The requirement for two interaction regions is interpreted in the regulatory framework of a substrate-dependent conformational change that brings those two regions into an orientation optimal for binding IIAGlc.