Yao Yong, Ding Yi, Tian Ye, Opella Stanley J, Marassi Francesca M
Sanford Burnham Medical Research Institute, La Jolla, CA, USA.
Methods Mol Biol. 2013;1063:145-58. doi: 10.1007/978-1-62703-583-5_8.
NMR spectroscopy enables the structures of membrane proteins to be determined in the native-like environment of the phospholipid bilayer membrane. This chapter outlines the methods for membrane protein structural studies using solid-state NMR spectroscopy with samples of membrane proteins incorporated in proteoliposomes or planar lipid bilayers. The methods for protein expression and purification, sample preparation, and NMR experiments are described and illustrated with examples from OmpX and Ail, two bacterial outer membrane proteins that function in bacterial virulence.
核磁共振光谱法能够在磷脂双分子层膜的类天然环境中确定膜蛋白的结构。本章概述了使用固态核磁共振光谱法对膜蛋白进行结构研究的方法,这些样品是掺入蛋白脂质体或平面脂质双分子层中的膜蛋白。文中描述了蛋白质表达与纯化、样品制备及核磁共振实验的方法,并以OmpX和Ail这两种在细菌致病性中起作用的细菌外膜蛋白为例进行说明。