Luciferase complementation based-detection of G-protein-coupled receptor activity. (17th July 2018)

• Record Type: Journal Article
• Authors: Yano, Hideaki; Cai, Ning Sheng; Javitch, Jonathan A; Ferré, Sergi
• Is Part Of: Biotechniques. Volume 65:Number 1(2018)
• Page Start: 9
• Page End: 14
• Publication Date: 2018-07-17
• Subjects: assay development -- biosensor -- complementation -- dopamine receptor -- GPCR -- luciferase; Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/btn-2018-0039
• Languages: English
• ISSNs: 0736-6205

Abstract

Protein complementation assays (PCA) are used as pharmacological tools, enabling a wide array of applications, ranging from studies of protein–protein interactions to second messenger effects. Methods to detect activities of G protein-coupled receptors (GPCRs) have particular relevance for drug screening. Recent development of an engineered luciferase NanoLuc created the possibility of generating a novel PCA, which in turn could open a new avenue for developing drug screening assays. Here we identified a novel split position for NanoLuc and demonstrated its use in a series of fusion constructs to detect the activity of GPCRs. The split construct can be applied to a variety of pharmacological screening systems.