title: Functional expression and characterization of the C. elegans G-protein-coupled FLP-2 Receptor (T19F4.1) in mammalian cells and yeast.
abstract: Profound neuropeptide diversity characterizes the nematode nervous system, but it has proven challenging to match neuropeptide G protein-coupled receptors (GPCR) with their cognate ligands in heterologous systems. We have expressed the Caenorhabditis elegans GPCR encoded in the locus T19F4.1, previously matched with FMRFamide-like peptides encoded on the flp-2 precursor gene, in mammalian cells and in the yeast Saccharomyces cerevisiae. Pharmacological characterization revealed that the receptor is potently activated by flp-2 peptides in CHO cells (∼10 nM EC50) and in yeast (∼100 nM EC50), signaling through a Gqα pathway in each system. The yeast GPCR expression system provides a robust assay for screening for agonists of the flp-2 receptor and is the target of an ongoing high-throughput screening exercise.
authors.affiliation: Pfizer Animal Health Discovery Research, Veterinary Medicine Discovery Research, Kalamazoo, MI 49001, USA.
authors.forename: Martha J
authors.initials: MJ
authors.lastname: Larsen
authors.affiliation: Institute of Parasitology, McGill University, 21,111 Lakeshore Road, Ste-Anne de Bellevue, Quebec, Canada H9X 3V9.
authors.forename: Elizabeth Ruiz
authors.initials: ER
authors.lastname: Lancheros
authors.affiliation: Pfizer Animal Health Discovery Research, Veterinary Medicine Discovery Research, Kalamazoo, MI 49001, USA.
authors.forename: Tracey
authors.initials: T
authors.lastname: Williams
authors.affiliation: Pfizer Animal Health Discovery Research, Veterinary Medicine Discovery Research, Kalamazoo, MI 49001, USA.
authors.forename: David E
authors.initials: DE
authors.lastname: Lowery
authors.affiliation: Pfizer Animal Health Discovery Research, Veterinary Medicine Discovery Research, Kalamazoo, MI 49001, USA.
authors.forename: Timothy G
authors.initials: TG
authors.lastname: Geary
authors.affiliation: Pfizer Animal Health Discovery Research, Veterinary Medicine Discovery Research, Kalamazoo, MI 49001, USA.
authors.forename: Teresa M
authors.initials: TM
authors.lastname: Kubiak
keywords: Caenorhabditis elegans; FLIPR, fluorescence imaging plate reader; FLP; FLP, FMRFamide-like peptide; FLP2-R, receptor for peptides encoded on flp-2; GPCR, G protein-coupled receptor; Neuropeptide; flp, FMRFamide-like protein gene; flp2; nlp, neuropeptide-like protein gene