A cell-based protein-protein interaction method using a permuted luciferase reporter.

Current chemical genomics Pub Date : 2011-01-01 Epub Date: 2011-11-30 DOI:10.2174/1875397301105010122
Haifeng Eishingdrelo, Jidong Cai, Paul Weissensee, Praveen Sharma, Michael J Tocci, Paul S Wright
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引用次数: 20

Abstract

We have developed a novel cell-based protein-protein interaction assay method. The method relies on conversion of an inactive permuted luciferase containing a Tobacco Etch Virus protease (TEV) cleavage sequence fused onto protein (A) to an active luciferase upon interaction and cleavage by another protein (B) fused with the TEV protease. We demonstrate assay applicability for ligand-induced protein-protein interactions including G-protein coupled receptors, receptor tyrosine kinases and nuclear hormone receptors.

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一种基于细胞的蛋白-蛋白相互作用方法,使用排列荧光素酶报告基因。
我们开发了一种新的基于细胞的蛋白质-蛋白质相互作用测定方法。该方法依赖于含有融合到蛋白质(a)上的烟草蚀刻病毒蛋白酶(TEV)切割序列的无活性排列荧光素酶在与TEV蛋白酶融合的另一蛋白质(B)相互作用和切割后转化为活性荧光素酶。我们证明了该方法适用于配体诱导的蛋白质相互作用,包括g蛋白偶联受体、酪氨酸激酶受体和核激素受体。
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