Zhou Yizhou, Blanco Luz P, Smith Daniel R, Chapman Matthew R
Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI, USA.
Methods Mol Biol. 2012;849:303-20. doi: 10.1007/978-1-61779-551-0_21.
Many bacteria can assemble functional amyloid fibers on their cell surface. The majority of bacterial amyloids contribute to biofilm or other community behaviors where cells interact with a surface or with another cell. Bacterial amyloids, like all functional amyloids, share structural and biochemical properties with disease-associated eukaryotic amyloids. The general ability of amyloids to bind amyloid-specific dyes, such as Congo red, and their resistance to denaturation have provided useful tools for scoring and quantifying bacterial amyloid formation. Here, we present basic approaches to study bacterial amyloids by focusing on the well-studied curli amyloid fibers expressed by Enterobacteriaceae. These methods exploit the specific tinctorial and biophysical properties of amyloids. The methods described here are straightforward and can be easily applied by any modern molecular biology lab for the study of other bacterial amyloids.
许多细菌能够在其细胞表面组装功能性淀粉样纤维。大多数细菌淀粉样蛋白有助于生物膜或其他群体行为,在这些行为中细胞与表面或其他细胞相互作用。细菌淀粉样蛋白与所有功能性淀粉样蛋白一样,与疾病相关的真核淀粉样蛋白具有结构和生化特性。淀粉样蛋白结合淀粉样特异性染料(如刚果红)的一般能力及其对变性的抗性,为评估和量化细菌淀粉样蛋白的形成提供了有用的工具。在这里,我们通过关注肠杆菌科表达的研究充分的卷曲淀粉样纤维,介绍研究细菌淀粉样蛋白的基本方法。这些方法利用了淀粉样蛋白的特定染色和生物物理特性。这里描述的方法很简单,任何现代分子生物学实验室都可以很容易地应用于研究其他细菌淀粉样蛋白。