Locating a Protein−Protein Interaction in Living Cells via Split Renilla Luciferase Complementation
- 15 July 2003
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 75 (16), 4176-4181
- https://doi.org/10.1021/ac0300800
Abstract
For spatial and quantitative kinetic analysis of protein−protein interactions (PPIs) in living mammalian cells, a method was developed in which PPI-induced complementation of split Renilla luciferase triggers spontaneous emission of luminescence using a cell membrane permeable substrate, coelenterazine. This split Renilla luciferase complementation readout was shown to work for locating a PPI between the tyrosine-phosphorylated peptide (Y941) of IRS-1 and the SH2 domain of PI3K among insulin signaling pathways in living Chinese hamster ovary cells overexpressing human insulin receptors (CHO-HIR). It was thereby found that the insulin-stimulated interaction occurred near the plasma membrane in the cytosol.This publication has 14 references indexed in Scilit:
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