使用基于HaloTag的荧光传感器监测活细胞中的蛋白质组应激。
Monitoring Proteome Stress in Live Cells Using HaloTag-Based Fluorogenic Sensor.
作者信息
Liu Yu, Fares Matthew, Zhang Xin
机构信息
Department of Chemistry, The Pennsylvania State University, University Park, PA, USA.
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, USA.
出版信息
Methods Mol Biol. 2019;1873:171-182. doi: 10.1007/978-1-4939-8820-4_10.
Fluorescent folding sensor is a powerful tool to detect proteome stresses, including heat, osmotic, oxidative, and drug induced stresses. Monitoring proteome stress using these sensors allows us to dissect the mechanism of cellular stress and find therapeutics that ameliorate stress related diseases. Here we present a HaloTag-based fluorogenic proteome stress sensor (AgHalo) to robustly detect and quantify proteome stresses in live cells. We describe how proteome stresses are monitored in both bacterial and mammalian live cells using fluorescence confocal microscope and fluorescence plate reader.
荧光折叠传感器是检测蛋白质组应激的有力工具，包括热应激、渗透应激、氧化应激和药物诱导的应激。使用这些传感器监测蛋白质组应激使我们能够剖析细胞应激的机制，并找到改善应激相关疾病的治疗方法。在这里，我们展示了一种基于卤代标签的荧光蛋白质组应激传感器（AgHalo），用于在活细胞中稳健地检测和量化蛋白质组应激。我们描述了如何使用荧光共聚焦显微镜和荧光酶标仪在细菌和哺乳动物活细胞中监测蛋白质组应激。
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