Hankins Jessica V, Madsen James A, Needham Brittany D, Brodbelt Jennifer S, Trent M Stephen
Section of Molecular Genetics and Microbiology, The University of Texas at Austin, Austin, TX 78712, USA.
Department of Chemistry and Biochemistry, The University of Texas at Austin, Austin, TX 78712, USA.
Methods Mol Biol. 2013;966:239-258. doi: 10.1007/978-1-62703-245-2_15.
The isolation and characterization of the lipid A domain of lipopolysaccharide (LPS) are important methodologies utilized to gain understanding of the Gram-negative cell envelope. Here, we describe protocols often employed by our laboratory for small- and large-scale isolation of lipid A from bacterial cells. Additionally, we describe various methodologies including isolation of radiolabeled lipid A, thin layer chromatography, and various mass spectrometry methods. Tandem mass spectrometry is an integral tool for the structural characterization of lipid A molecules, and both coventional collision induced dissociation (CID) and new ultraviolet photodissociation (UVPD) methods are described.
脂多糖（LPS）脂质A结构域的分离与表征是用于深入了解革兰氏阴性菌细胞壁的重要方法。在此，我们描述了本实验室常用于从细菌细胞中进行小规模和大规模脂质A分离的方案。此外，我们还描述了各种方法，包括放射性标记脂质A的分离、薄层色谱法以及各种质谱方法。串联质谱是脂质A分子结构表征的重要工具，本文将同时介绍传统的碰撞诱导解离（CID）方法和新的紫外光解离（UVPD）方法。
