Department of Microbiology, Sapporo Medical University School of Medicine, Sapporo, Japan.
Methods Mol Biol. 2024;2851:39-60. doi: 10.1007/978-1-0716-4096-8_4.
Gram-positive bacteria, including lactic acid bacteria (LAB), possess lipoteichoic acid (LTA) on the cell surface. LTA is an amphiphilic molecule typically composed of hydrophilic glycerolphosphate polymer and hydrophobic anchor glycolipid moieties. It is involved in physiological properties of the cell surface and also plays roles in interactions with the host. Appropriate preparation procedures, such as extraction and purification, are required to clarify the structure-activity relationship. Structural diversity of LTA has been reported at the bacterial species and strain levels, and structural differences might affect interactions with the host. This chapter introduces techniques for preparation and structural analysis of LTA derived from LAB. It consists of four sections, covering butanol extraction, hydrophobic interaction chromatography, immunoblotting, and structural analysis. Technical notes containing supplemental information about the individual steps are also provided.
革兰氏阳性菌,包括乳酸菌 (LAB),在细胞表面具有脂磷壁酸 (LTA)。LTA 是一种两亲性分子,通常由亲水性甘油磷酸聚合物和疏水性锚定糖脂部分组成。它参与细胞表面的生理特性,也在与宿主的相互作用中发挥作用。需要适当的准备程序,如提取和纯化,以阐明结构-活性关系。在细菌种和菌株水平上已经报道了 LTA 的结构多样性,结构差异可能影响与宿主的相互作用。本章介绍了从 LAB 中提取和分析 LTA 的结构的技术。它包括四个部分,涵盖丁醇提取、疏水相互作用色谱、免疫印迹和结构分析。还提供了关于各个步骤的补充信息的技术说明。