Identification of surrogate agonists for the human FPRL-1 receptor by autocrine selection in yeast

• Published in: Nature biotechnology Vol. 16; no. 13; pp. 1334 - 1337
• Main Authors: Klein, Christine, Paul, Jeremy I, Sauvé, Karen, Schmidt, Mary M, Arcangeli, Loretta, Ransom, John, Trueheart, Joshua, Manfredi, John P, Broach, James R, Murphy, Andrew J
• Format: Journal Article
• Language: English
• Published: New York, NY Nature 01.12.1998
• Subjects: Amino Acid Sequence; Biological and medical sciences; Cell Line; Diverse techniques; Fundamental and applied biological sciences. Psychology; Humans; Ligands; Molecular and cellular biology; Molecular Sequence Data; Monocytes - metabolism; N-Formylmethionine Leucyl-Phenylalanine - metabolism; Neutrophils - metabolism; Peptides - chemistry; Peptides - metabolism; Receptors, Formyl Peptide; Receptors, Immunologic - agonists; Receptors, Immunologic - metabolism; Receptors, Peptide - agonists; Receptors, Peptide - metabolism; Recombinant Proteins - agonists; Recombinant Proteins - metabolism; Saccharomyces cerevisiae - genetics; Fungi; Recombinant microorganism; Signal transduction; Agonist; Yeast; Ascomycetes; Pharmacology; Method; Saccharomyces cerevisiae; Orphan receptor; Thallophyta; G protein
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/4310

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.