Departament de Fisicoquimica, Facultat de Farmacia, and Institut de Nanociencia i Nanotecnologia (IN2UB), and Laboratori de Quimica Farmaceutica (Unitat Associada al CSIC), Facultat de Farmacia, and Institut de Biomedicina (IBUB), Universitat de Barcelona, Av. Joan XXIII, 27-31, 08028-Barcelona, Spain.
Curr Med Chem. 2014;21(9):1152-9. doi: 10.2174/09298673113206660256.
Amyloid aggregation is linked to a large number of human disorders, from neurodegenerative diseases as Alzheimer's disease (AD) or spongiform encephalopathies to non-neuropathic localized diseases as type II diabetes and cataracts. Because the formation of insoluble inclusion bodies (IBs) during recombinant protein production in bacteria has been recently shown to share mechanistic features with amyloid self-assembly, bacteria have emerged as a tool to study amyloid aggregation. Herein we present a fast, simple, inexpensive and quantitative method for the screening of potential anti-aggregating drugs. This method is based on monitoring the changes in the binding of thioflavin-S to intracellular IBs in intact Eschericchia coli cells in the presence of small chemical compounds. This in vivo technique fairly recapitulates previous in vitro data. Here we mainly use the Alzheimer's related β-amyloid peptide as a model system, but the technique can be easily implemented for screening inhibitors relevant for other conformational diseases simply by changing the recombinant amyloid protein target. Indeed, we show that this methodology can be also applied to the evaluation of inhibitors of the aggregation of tau protein, another amyloidogenic protein with a key role in AD.
淀粉样蛋白聚集与许多人类疾病有关，从神经退行性疾病如阿尔茨海默病（AD）或海绵状脑病到非神经病变的局部疾病如 2 型糖尿病和白内障。由于最近已经表明细菌中重组蛋白生产过程中不可溶包涵体（IBs）的形成与淀粉样蛋白自组装具有机制特征，因此细菌已成为研究淀粉样蛋白聚集的工具。在此，我们提出了一种快速、简单、廉价且定量的方法，用于筛选潜在的抗聚集药物。该方法基于监测小分子化合物存在时完整大肠埃希氏菌细胞内 IBs 与硫黄素-S 结合的变化。这种体内技术很好地重现了以前的体外数据。在这里，我们主要使用与阿尔茨海默病相关的β-淀粉样肽作为模型系统，但该技术可以通过简单地改变重组淀粉样蛋白靶标，很容易地用于筛选与其他构象疾病相关的抑制剂。事实上，我们表明该方法也可以应用于评估 tau 蛋白聚集抑制剂，tau 蛋白是另一种在 AD 中起关键作用的淀粉样蛋白。
