Abstract
We describe a procedure for isolating agonists for mammalian G protein–coupled receptors of unknown function. Human formyl peptide receptor like-1 (FPRL-1) receptor, originally identified as an orphan G protein–coupled receptor related to the formyl peptide receptor (FPR1), was expressed in Saccharomyces cells designed to couple receptor activation to histidine prototrophy. Selection for histidine prototrophs among transformants obtained with a plasmid-based library encoding random peptides identified six different agonists, each of whose production yielded autocrine stimulation of the receptor expressed in yeast. A synthetic version of each peptide promoted activation of FPRL-1 expressed in human embryonic kidney (HEK293) cells, and five of the peptides exhibited significant selectivity for activation of FPRL-1 relative to FPR1. One selective peptide was tested and found to mobilize calcium in isolated human neutrophils. This demonstrates that stimulation of FPRL-1 results in neutrophil activation and suggests that the receptor functions as a component of the inflammatory response. This autocrine selection protocol may be a generally applicable method for providing pharmacological tools to evaluate the physiological roles of the growing number of mammalian orphan G protein–coupled receptors.
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Acknowledgements
We thank Philip Murphy for providing plasmids encoding FPR and FPRL-1; William T. Wiesler for Cp1584 and β-galactosidase assays; Anna Farescal and Dwight Morrow for conducting peptide searches; Martin McKinney and Lorna Nuñez for technical assistance; and Nan-Xin Qian for providing the HEK293 cell line expressing Gα16. We also thank David R. Webb for a critical reading of the manuscript.
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Klein, C., Paul, J., Sauvé, K. et al. Identification of surrogate agonists for the human FPRL-1 receptor by autocrine selection in yeast. Nat Biotechnol 16, 1334–1337 (1998). https://doi.org/10.1038/4310
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DOI: https://doi.org/10.1038/4310
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