Zhou Yizhou, Smith Daniel R, Hufnagel David A, Chapman Matthew R
Department of Molecular, Cellular, and Development Biology, University of Michigan, Ann Arbor, MI, USA.
Methods Mol Biol. 2013;966:53-75. doi: 10.1007/978-1-62703-245-2_4.
Curli are proteinaceous fibrous structures produced on the surface of many gram-negative bacteria. As a major constituent of the extracellular matrix, curli mediate interactions between the bacteria and its environment, and as such, curli play a critical role in biofilm formation. Curli fibers share biophysical properties with a growing number of remarkably stable and ordered protein aggregates called amyloid. Here we describe experimental methods to study the biogenesis and assembly of curli by exploiting their amyloid properties. We also present methods to analyze curli-mediated biofilm formation. These approaches are straightforward and can easily be adapted to study other bacterially produced amyloids.
卷曲菌毛是在许多革兰氏阴性菌表面产生的蛋白质纤维结构。作为细胞外基质的主要成分,卷曲菌毛介导细菌与其环境之间的相互作用,因此,卷曲菌毛在生物膜形成中起关键作用。卷曲菌毛纤维与越来越多被称为淀粉样蛋白的极其稳定且有序的蛋白质聚集体具有共同的生物物理特性。在此,我们描述了利用卷曲菌毛的淀粉样特性来研究其生物发生和组装的实验方法。我们还介绍了分析卷曲菌毛介导的生物膜形成的方法。这些方法很直接,并且可以很容易地适用于研究其他细菌产生的淀粉样蛋白。