Vadukul Devkee M, Al-Hilaly Youssra K, Serpell Louise C
School of Life Sciences, University of Sussex, East Sussex, UK.
Department of Chemistry, College of Sciences, Al-Mustansiriyah University, Baghdad, Iraq.
Methods Mol Biol. 2019;1873:109-122. doi: 10.1007/978-1-4939-8820-4_7.
Many proteins and peptides are able to self-assemble in solution in vitro and in vivo to form amyloid-like fibrils. These fibrils share common structural characteristics. In order for a fibril to be characterized as amyloid, it is expected to fit certain criteria including the composition of cross-β. Here we describe how the formation of amyloid fibrils can be characterized in vitro using a variety of methods including circular dichroism and intrinsic tyrosine/tryptophan fluoresence to follow conformational changes; Thioflavin and/or ThS assembly to monitor nucleation and growth; transmission electron microscopy to visualize fibrillar morphology and X-ray fiber diffraction to examine cross-β structure.
许多蛋白质和肽能够在体外溶液以及体内自组装形成淀粉样纤维。这些纤维具有共同的结构特征。为了将一种纤维表征为淀粉样蛋白，预期它要符合某些标准，包括交叉β结构的组成。在这里，我们描述了如何在体外使用多种方法来表征淀粉样纤维的形成，这些方法包括利用圆二色性和内在酪氨酸/色氨酸荧光来跟踪构象变化；硫黄素和/或硫黄素S组装以监测成核和生长；透射电子显微镜观察纤维形态以及X射线纤维衍射检查交叉β结构。
