Development and characterization of a potent free fatty acid receptor 1 (FFA1) fluorescent tracer

Christiansen, E., Hudson, B. D. , Hansen, A. H., Milligan, G. and Ulven, T. (2016) Development and characterization of a potent free fatty acid receptor 1 (FFA1) fluorescent tracer. Journal of Medicinal Chemistry, 59(10), pp. 4849-4858. (doi: 10.1021/acs.jmedchem.6b00202) (PMID:27074625)

118427.pdf - Accepted Version



The free fatty acid receptor 1 (FFA1/GPR40) is a potential target for treatment of type 2 diabetes. Although several potent agonists have been described, there remains a strong need for suitable tracers to interrogate ligand binding to this receptor. We address this by exploring fluorophore-tethering to known potent FFA1 agonists. This led to the development of 4, a high affinity FFA1 tracer with favorable and polarity-dependent fluorescent properties. A close to ideal overlap between the emission spectrum of the NanoLuciferase receptor tag and the excitation spectrum of 4 enabled the establishment of a homogenous BRET-based binding assay suitable for both detailed kinetic studies and high throughput competition binding studies. Using 4 as a tracer demonstrated that the compound acts fully competitively with selected synthetic agonists but not with lauric acid and allowed for the characterization of binding affinities of a diverse selection of known FFA1 agonists, indicating that 4 will be a valuable tool for future studies at FFA1.

Item Type:Articles
Additional Information:The study has been funded by the Danish Council for Independent Research | Technology and Production Sciences (grant 09-070364), the Danish Council for Strategic Research (grant 11-116196) and the University of Southern Denmark. EC is supported by a fellowship from the Lundbeck Foundation. BDH is supported by a fellowship from the University of Glasgow.
Glasgow Author(s) Enlighten ID:Hudson, Dr Brian and Milligan, Professor Graeme
Authors: Christiansen, E., Hudson, B. D., Hansen, A. H., Milligan, G., and Ulven, T.
College/School:College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Journal Name:Journal of Medicinal Chemistry
Publisher:American Chemical Society
ISSN (Online):1520-4804
Published Online:13 April 2016
Copyright Holders:Copyright © 2016 American Chemical Society
First Published:First published in Journal of Medicinal Chemistry 59(10):4849-4858
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher

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