乙酰胆碱酯酶抑制分析用于高通量筛选。
Acetylcholinesterase Inhibition Assays for High-Throughput Screening.
机构信息
National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
Division of Toxicology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration, Laurel, MD, USA.
出版信息
Methods Mol Biol. 2022;2474:47-58. doi: 10.1007/978-1-0716-2213-1_6.
Acetylcholinesterase (AChE) hydrolyzes acetylcholine (ACh), a vital neurotransmitter that regulates muscle movement and brain function, including memory, attention, and learning. Inhibition of AChE activity can cause a variety of adverse health effects and toxicity. Identifying AChE inhibitors quickly and efficiently warrants developing AChE inhibition assays in a quantitative, high-throughput screening (qHTS) platform. In this chapter, protocols for multiple homogenous AChE inhibition assays used in a qHTS system are provided. These AChE inhibition assays include a (1) human neuroblastoma (SH-SY5Y) cell-based assay with fluorescence or colorimetric detection; (2) human recombinant AChE with fluorescence or colorimetric detection; and (3) combination of human recombinant AChE and liver microsomes with colorimetric detection, which enables detection of test compounds requiring metabolic activation to become AChE inhibitors. Together, these AChE assays can help identify, prioritize, and predict chemical hazards in large compound libraries using qHTS systems.
乙酰胆碱酯酶 (AChE) 可水解乙酰胆碱 (ACh)，后者是一种重要的神经递质，可调节肌肉运动和大脑功能，包括记忆、注意力和学习。AChE 活性的抑制可能会导致各种健康不良影响和毒性。因此，快速有效地鉴定 AChE 抑制剂需要在定量、高通量筛选 (qHTS) 平台中开发 AChE 抑制测定法。本章提供了在 qHTS 系统中使用的多种均相 AChE 抑制测定法的方案。这些 AChE 抑制测定法包括：(1) 具有荧光或比色检测的人神经母细胞瘤 (SH-SY5Y) 细胞测定法；(2) 具有荧光或比色检测的人重组 AChE；和 (3) 人重组 AChE 和肝微粒体的组合测定法，具有比色检测，可检测需要代谢激活才能成为 AChE 抑制剂的测试化合物。总之，这些 AChE 测定法可帮助使用 qHTS 系统在大型化合物文库中识别、优先考虑和预测化学危害。
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