Hatters Danny M, Griffin Michael D W
Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, VIC, Australia.
Methods Mol Biol. 2011;752:121-36. doi: 10.1007/978-1-60327-223-0_8.
Twenty-five proteins are known to form amyloid fibrils in vivo in association with disease (Westermark et al., Amyloid 12:1-4, 2005). However, the fundamental ability of a protein to form amyloid-like fibrils is far more widespread than in just the proteins associated with disease, and indeed this property can provide insight into the basic thermodynamics of folding and misfolding pathways. But how does one determine whether a protein has formed amyloid-like fibrils? In this chapter, we cover the basic steps toward defining the amyloid-like properties of a protein and how to measure the kinetics of fibrillization. We describe several basic tests for aggregation and the binding to two classic amyloid-reactive dyes, Congo Red, and thioflavin T, which are key indicators to the presence of fibrils.
已知有25种蛋白质在体内与疾病相关联形成淀粉样纤维（韦斯特马克等人，《淀粉样蛋白》12:1 - 4，2005年）。然而，蛋白质形成淀粉样纤维的基本能力远比仅与疾病相关的蛋白质更为广泛，实际上，这一特性可以为折叠和错误折叠途径的基本热力学提供见解。但是如何确定一种蛋白质是否形成了淀粉样纤维呢？在本章中，我们涵盖了定义蛋白质淀粉样特性的基本步骤以及如何测量纤维化动力学。我们描述了几种用于聚集以及与两种经典淀粉样反应染料（刚果红和硫黄素T）结合的基本测试，这两种染料是纤维存在的关键指标。
