The outer membrane of Gram-negative bacteria: lipid A isolation and characterization.
The outer membrane of Gram-negative bacteria: lipid A isolation and characterization.
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DOI：
10.1007/978-1-62703-245-2_15
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Trent MS
中科院分区：
文献类型：
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作者：
Hankins JV;Madsen JA;Needham BD;Brodbelt JS;Trent MS
脂多糖的脂类A结构域的分离和鉴定是了解革兰氏阴性细胞被膜的重要方法。在这里，我们描述了我们实验室经常使用的从细菌细胞中小规模和大规模分离类脂A的方法。此外，我们还介绍了各种方法，包括放射性标记脂类A的分离、薄层层析和各种质谱学方法。串联质谱仪是表征脂类A分子结构的重要工具，介绍了传统的碰撞诱导解离(CID)和新的紫外光解离(UVPD)方法。
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.