Identification of surrogate agonists for the human FPRL-1 receptor by autocrine selection in yeast
| Autor: | Loretta Arcangeli, Mary M. Schmidt, Karen Sauvé, James R. Broach, John Ransom, Andrew J. Murphy, Christine Klein, Jeremy Paul, Joshua Trueheart, John Manfredi |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Receptors
Peptide Neutrophils Molecular Sequence Data Biomedical Engineering Bioengineering Saccharomyces cerevisiae Biology Ligands Applied Microbiology and Biotechnology Monocytes Cell Line Enzyme-linked receptor Humans 5-HT5A receptor Amino Acid Sequence Receptors Immunologic Glucagon-like peptide 1 receptor Orphan receptor Formyl peptide receptor Receptors Formyl Peptide Recombinant Proteins Neuron-derived orphan receptor 1 N-Formylmethionine Leucyl-Phenylalanine Biochemistry Interleukin-21 receptor Molecular Medicine Estrogen-related receptor gamma Peptides Biotechnology |
| Zdroj: | Nature biotechnology. 16(13) |
| ISSN: | 1087-0156 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|