A luminescent assay for real-time measurements of receptor endocytosis in living cells
A luminescent assay for real-time measurements of receptor endocytosis in living cells
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DOI：
10.1016/j.ab.2015.08.005
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发表时间：
2015-11-15
影响因子：
2.9
通讯作者：
Wood, Keith V.
中科院分区：
文献类型：
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作者：
Robers, Matthew B.;Binkowski, Brock F.;Wood, Keith V.
配体介导的内吞作用是控制细胞表面受体发出信号的持续时间和强度的关键自调节机制。由于内吞作用的机制复杂性及其在疾病中的新兴相关性，能够在细胞中跟踪这一动态过程的简单方法变得越来越需要。我们已经开发了一种生物发光报告技术，用于实时分析配体介导的受体内吞作用，使用NanoLuc荧光素酶与各种g蛋白偶联受体（gpcr）的遗传融合。该方法与标准微孔板格式兼容，这将减少高通量筛选的工作流程。本文还介绍了该技术在表皮生长因子受体（EGFR）内吞作用中的应用，证明了该方法在gpcr之外的潜在适用性。(C) 2015爱思唯尔公司版权所有。
Ligand-mediated endocytosis is a key autoregulatory mechanism governing the duration and intensity of signals emanating from cell surface receptors. Due to the mechanistic complexity of endocytosis and its emerging relevance in disease, simple methods capable of tracking this dynamic process in cells have become increasingly desirable. We have developed a bioluminescent reporter technology for real-time analysis of ligand-mediated receptor endocytosis using genetic fusions of NanoLuc luciferase with various G-protein-coupled receptors (GPCRs). This method is compatible with standard microplate formats, which should decrease work flows for high-throughput screens. This article also describes the application of this technology to endocytosis of epidermal growth factor receptor (EGFR), demonstrating potential applicability of the method beyond GPCRs. (C) 2015 Elsevier Inc. All rights reserved.