Centro de Genómica y Bioinformática, Facultad de Ciencias, Universidad Mayor, Santiago, Chile.
Methods Mol Biol. 2020;2136:347-364. doi: 10.1007/978-1-0716-0467-0_28.
Gram-positive bacteria use their adhesive pili to attach to host cells during early stages of a bacterial infection. These extracellular hair-like appendages experience mechanical stresses of hundreds of picoNewtons; however, the presence of an internal isopeptide bond prevents the pilus protein from unfolding. Here, we describe a method to interfere with nascent pili proteins through a peptide that mimics one of the β-strands of the molecule. By using AFM-based force spectroscopy, we study the isopeptide bond formation and the effect of the peptide in the elasticity of the pilus protein. This method could be used to afford a new strategy for mechanically targeted antibiotics by simply blocking the folding of the bacterial pilus protein.
革兰氏阳性菌在细菌感染的早期阶段使用其黏附性菌毛附着在宿主细胞上。这些细胞外的毛发状附属物经历数百皮牛顿的机械应力；然而，内部异肽键的存在阻止了菌毛蛋白的展开。在这里，我们描述了一种通过模拟分子中β-链之一的肽来干扰新生菌毛蛋白的方法。通过使用基于 AFM 的力谱法，我们研究了异肽键的形成以及肽对菌毛蛋白弹性的影响。这种方法可用于通过简单地阻止细菌菌毛蛋白的折叠来提供一种针对机械靶向抗生素的新策略。
