利用双分子发光互补(BiLC)可视化活细胞中蛋白质相互作用

Q1 Biochemistry, Genetics and Molecular Biology
Lisette G.G.C. Verhoef, Mark Wade
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引用次数: 3

摘要

细胞内蛋白质-蛋白质相互作用(PPIs)的数量远远超过基因组编码的蛋白质总数。动态细胞PPI网络响应外部刺激和内源性代谢,以维持体内平衡。许多ppi直接参与疾病发病机制和/或对治疗药物的耐药性;因此,它们代表了潜在的药物靶点。一种通常被称为“双分子互补”的技术依赖于一个分子报告蛋白(如荧光蛋白或发光蛋白)的物理分裂,并将产生的两个片段融合成一对相互作用的蛋白。当这些蛋白质相互作用时,它们有效地重建了分子报告蛋白的活性(通常导致荧光或发光增加)。本单元描述了用于报告细胞内ppi的双分子发光互补(BiLC)测定的选择和发展,并提供了BiLC用于识别可以调节ppi的小分子的示例。©2017 by John Wiley &儿子,Inc。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Visualization of Protein Interactions in Living Cells Using Bimolecular Luminescence Complementation (BiLC)

The number of intracellular protein-protein interactions (PPIs) far exceeds the total number of proteins encoded by the genome. Dynamic cellular PPI networks respond to external stimuli and endogenous metabolism in order to maintain homeostasis. Many PPIs are directly involved in disease pathogenesis and/or resistance to therapeutics; they therefore represent potential drug targets. A technology generally termed ‘bimolecular complementation’ relies on the physical splitting of a molecular reporter (such as a fluorescent or luminescent protein) and fusion of the resulting two fragments to a pair of interacting proteins. When these proteins interact, they effectively reconstitute the activity of the molecular reporter (typically leading to increased fluorescence or luminescence). This unit describes the selection and development of bimolecular luminescence complementation (BiLC) assays for reporting intracellular PPIs, and provides examples in which BiLC was used to identify small molecules that can modulate PPIs. © 2017 by John Wiley & Sons, Inc.

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来源期刊
Current Protocols in Protein Science
Current Protocols in Protein Science Biochemistry, Genetics and Molecular Biology-Biochemistry
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期刊介绍: With the mapping of the human genome, more and more researchers are exploring protein structures and functions in living organisms. Current Protocols in Protein Science provides protein scientists, biochemists, molecular biologists, geneticists, and others with the first comprehensive suite of protocols for this growing field.
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