Noinaj Nicholas, Kuszak Adam J, Buchanan Susan K
Department of Biological Sciences, Markey Center for Structural Biology, Purdue University, West Lafayette, IN, 47907, USA.
Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, 20892, USA.
Methods Mol Biol. 2015;1329:51-6. doi: 10.1007/978-1-4939-2871-2_4.
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.
β-桶状膜蛋白在某种程度上是独特的，因为它们的折叠状态可以通过半天然SDS-PAGE方法进行监测，以确定它们是否正确折叠。这种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完整细胞，以监测感兴趣蛋白质的折叠状态。此外，热可修饰性测定已被证明在研究BAM复合物及其在将β-桶状膜蛋白折叠并插入外膜中的作用方面不可或缺。在这里，我们描述了我们实验室在研究外膜蛋白时用于进行热可修饰性测定的方案。
