A luminescent assay for real-time measurements of receptor endocytosis in living cells.

Autor: Robers MB; Promega Corporation, Fitchburg, WI 53711, USA. Electronic address: matt.robers@promega.com., Binkowski BF; Promega Corporation, Fitchburg, WI 53711, USA., Cong M; Promega Corporation, Fitchburg, WI 53711, USA., Zimprich C; Promega Corporation, Fitchburg, WI 53711, USA., Corona C; Promega Biosciences, San Luis Obispo, CA 93401, USA., McDougall M; Promega Biosciences, San Luis Obispo, CA 93401, USA., Otto G; Promega Corporation, Fitchburg, WI 53711, USA., Eggers CT; Promega Corporation, Fitchburg, WI 53711, USA., Hartnett J; Promega Corporation, Fitchburg, WI 53711, USA., Machleidt T; Promega Corporation, Fitchburg, WI 53711, USA., Fan F; Promega Corporation, Fitchburg, WI 53711, USA., Wood KV; Promega Corporation, Fitchburg, WI 53711, USA.
Jazyk: angličtina
Zdroj: Analytical biochemistry [Anal Biochem] 2015 Nov 15; Vol. 489, pp. 1-8. Date of Electronic Publication: 2015 Aug 13.
DOI: 10.1016/j.ab.2015.08.005
Abstrakt: Ligand-mediated endocytosis is a key autoregulatory mechanism governing the duration and intensity of signals emanating from cell surface receptors. Due to the mechanistic complexity of endocytosis and its emerging relevance in disease, simple methods capable of tracking this dynamic process in cells have become increasingly desirable. We have developed a bioluminescent reporter technology for real-time analysis of ligand-mediated receptor endocytosis using genetic fusions of NanoLuc luciferase with various G-protein-coupled receptors (GPCRs). This method is compatible with standard microplate formats, which should decrease work flows for high-throughput screens. This article also describes the application of this technology to endocytosis of epidermal growth factor receptor (EGFR), demonstrating potential applicability of the method beyond GPCRs.
(Copyright © 2015 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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