Heat Modifiability of Outer Membrane Proteins from Gram-Negative Bacteria.
Heat Modifiability of Outer Membrane Proteins from Gram-Negative Bacteria.
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DOI：
10.1007/978-1-4939-2871-2_4
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Buchanan SK
中科院分区：
文献类型：
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作者：
Noinaj N;Kuszak AJ;Buchanan SK
β-Barrel膜蛋白在某种程度上是独一无二的，因为可以使用半天然的SDSPAGE方法来监测它们的折叠状态，以确定它们是否正确折叠。这一特性，通常被称为热可修饰性，多年来一直用于纯化蛋白质和整个细胞，以监测目标蛋白质的折叠状态。此外，热可修饰性分析已被证明在研究BAM复合体及其在折叠和将β-Barrel膜蛋白插入外膜中的作用方面是必不可少的。在这里，我们描述了我们实验室在外膜蛋白研究中用于执行热可修饰性测试的方案。
β-barrel membrane proteins are somewhat unique in that their folding states can be monitored using semi-native SDS-PAGE methods to determine if they are folded properly or not. This property, which is commonly referred to as heat modifiability, has been used for many years on both purified protein and on whole cells to monitor folded states of proteins of interest. Additionally, heat modifiability assays have proven indispensable in studying the BAM complex and its role in folding and inserting β-barrel membrane proteins into the outer membrane. Here, we describe the protocol our lab uses for performing the heat modifiability assay in our studies on outer membrane proteins.