A rapid fluorogenic GPCR–β‐arrestin interaction assay |
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| Authors: | Qiang Zhang Yao‐Wu Zheng Shaun R Coughlin Xiaokun Shu |
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| Institution: | 1. Department of Pharmaceutical Chemistry, University of California—San Francisco, San Francisco, California;2. Cardiovascular Research Institute, University of California—San Francisco, San Francisco, California |
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| Abstract: | Detection of protein–protein interactions involved in signal transduction in live cells and organisms has a variety of important applications. We report a fluorogenic assay for G protein‐coupled receptor (GPCR)–β‐arrestin interaction that is genetically encoded, generalizes to multiple GPCRs, and features high signal‐to‐noise because fluorescence is absent until its components interact upon GPCR activation. Fluorescence after protease‐activated receptor‐1 activation developed in minutes and required specific serine–threonine residues in the receptor carboxyl tail, consistent with a classical G protein‐coupled receptor kinase dependent β‐arrestin recruitment mechanism. This assay provides a useful complement to other in vivo assays of GPCR activation. |
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| Keywords: | protein– protein interaction G protein coupled receptor fluorescent reporter green fluorescent protein |
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