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Development of a FLIPR Assay for the Simultaneous Identification of MrgD Agonists and Antagonists from a Single Screen
Journal article   Open access

Development of a FLIPR Assay for the Simultaneous Identification of MrgD Agonists and Antagonists from a Single Screen

Seena K Ajit, Mark H Pausch, Jeffrey D Kennedy and Edward J Kaftan
Journal of biomedicine & biotechnology, v 2010
2010
PMID: 20936132
url
https://doi.org/10.1155/2010/326020View
Published, Version of Record (VoR) Open

Abstract

MrgD, a member of the Mas-related gene family, is expressed exclusively in small diameter neurons in the dorsal root ganglion. This unique expression pattern, the presence of a single copy of MrgD in rodents and humans, and the identification of a putative ligand, beta-alanine, make it an experimentally attractive therapeutic target for pain with limited likelihood of side effects. We have devised a high throughput calcium mobilization assay that enables identification of both agonists and antagonists from a single screen for MrgD. Screening of the Library of Pharmacologically Active Compounds (LOPAC) validated this assay approach, and we identified both agonists and antagonists active at micromolar concentrations in MrgD expressing but not in parental CHO-DUKX cell line. Further characterization was performed using a subset of these screening hits. Our results demonstrated that the dual agonist/antagonist assay format is feasible and likely can be extended to most GPCRs with known agonist.

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11 citations in Scopus

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Collaboration types
Industry collaboration
Domestic collaboration
Web of Science research areas
Biotechnology & Applied Microbiology
Medicine, Research & Experimental
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