Li Gang, Peter Howard S
Department of Microbiology and Immunology, College of Medicine, University of Saskatchewan, Saskatoon, SK, Canada, S7N 5E5.
Methods Mol Biol. 2017;1615:143-149. doi: 10.1007/978-1-4939-7033-9_11.
Bacteria have developed a number of trans-envelope systems to transport molecules or assemble organelles across bacterial envelopes. However, bacterial envelopes contain a rigid netlike peptidoglycan structure that protects cells from osmotic lysis. Trans-envelope systems thus must interact with the peptidoglycan barrier to generate gaps or anchor structures to the peptidoglycan scaffold. Here we describe methods to use in vivo cross-linking and in vitro co-sedimentation to study protein-peptidoglycan interactions in Gram-negative bacteria. In particular, we address important considerations to ensure the specificity of the interactions in question.
细菌已经发展出多种跨膜系统来运输分子或在细菌包膜上组装细胞器。然而，细菌包膜含有一种刚性的网状肽聚糖结构，可保护细胞免受渗透裂解。因此，跨膜系统必须与肽聚糖屏障相互作用，以在肽聚糖支架上产生间隙或锚定结构。在这里，我们描述了使用体内交联和体外共沉降来研究革兰氏阴性菌中蛋白质-肽聚糖相互作用的方法。特别是，我们讨论了确保所研究相互作用特异性的重要注意事项。
