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Discovery of regulators of receptor internalization with high-throughput flow cytometry.

ABSTRACT: We developed a platform combining fluorogen-activating protein (FAP) technology with high-throughput flow cytometry to detect real-time protein trafficking to and from the plasma membrane in living cells. The hybrid platform facilitates drug discovery for trafficking receptors such as G protein-coupled receptors and was validated with the ??-adrenergic receptor (??AR) system. When a chemical library containing ?1200 off-patent drugs was screened against cells expressing FAP-tagged ??ARs, all 33 known ??AR-active ligands in the library were successfully identified, together with a number of compounds that might regulate receptor internalization in a nontraditional manner. Results indicated that the platform identified ligands of target proteins regardless of the associated signaling pathway; therefore, this approach presents opportunities to search for biased receptor modulators and is suitable for screening of multiplexed targets for improved efficiency. The results revealed that ligands may be biased with respect to the rate or duration of receptor internalization and that receptor internalization may be independent of activation of the mitogen-activated protein kinase pathway.

SUBMITTER: Wu Y

PROVIDER: S-EPMC3463215 | biostudies-literature | 2012 Oct

REPOSITORIES: biostudies-literature

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Discovery of regulators of receptor internalization with high-throughput flow cytometry.

Wu Yang Y Tapia Phillip H PH Fisher Gregory W GW Simons Peter C PC Strouse J Jacob JJ Foutz Terry T Waggoner Alan S AS Jarvik Jonathan J Sklar Larry A LA

Molecular pharmacology 20120705 4

We developed a platform combining fluorogen-activating protein (FAP) technology with high-throughput flow cytometry to detect real-time protein trafficking to and from the plasma membrane in living cells. The hybrid platform facilitates drug discovery for trafficking receptors such as G protein-coupled receptors and was validated with the β₂-adrenergic receptor (β₂AR) system. When a chemical library containing ∼1200 off-patent drugs was screened against cells expressing FAP-tagged β₂ARs, all 33 ...[more]

PMID: 22767611

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Dataset Information

Discovery of regulators of receptor internalization with high-throughput flow cytometry.

Publications

Discovery of regulators of receptor internalization with high-throughput flow cytometry.

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