Preparation of Caveolin-1 for NMR Spectroscopy Experiments.
Preparation of Caveolin-1 for NMR Spectroscopy Experiments.
复制标题
用于核磁共振波谱实验的 Caveolin-1 的制备。
DOI：
10.1007/978-1-0716-0732-9_13
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Glover,KerneyJebrell
中科院分区：
文献类型：
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作者：
Plucinsky,SarahM;Julien,JeffreyA;Glover,KerneyJebrell
Caveolin-1是一种20.5 kDa的完整膜蛋白，参与了多种细胞过程，包括信号转导、减轻细胞上的机械应力、内吞作用以及最重要的小窝形成。因此，人们对小窝蛋白1的结构特征产生了浓厚的兴趣。在众多可用的结构技术中，核磁共振波谱特别适合于研究具有显著的非结构化区域和不寻常的拓扑结构的完整的膜蛋白，如小窝蛋白-1。然而，该技术需要相对大量的蛋白质(即浓度在0.5-5 mm范围内)，而且获得这些量可能很困难，特别是对于像小窝蛋白-1这样的高度疏水的膜蛋白。在此，我们描述了一种稳健的制备小窝蛋白-1的方法，用于使用核磁共振进行结构研究。
Caveolin-1 is a 20.5 kDa integral membrane protein that is involved in a myriad of cellular processes including signal transduction, relieving mechano-stresses on the cell, endocytosis, and most importantly caveolae formation. As a consequence, there is intense interest in characterizing caveolin-1 structurally. Out of the many available structural techniques, nuclear magnetic resonance (NMR) spectroscopy is particularly well suited to investigations on integral membrane proteins like caveolin-1 that have significant unstructured regions and unusual topologies. However, the technique requires relatively large amounts of protein (i.e. concentrations in the 0.5–5 mM range), and obtaining these amounts can be difficult especially for highly hydrophobic membrane proteins such as caveolin-1. Herein, we describe a robust protocol for the preparation of caveolin-1 for structural studies using NMR.