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

Christine Klein, Jeremy I. Paul, Karen Sauvé, Mary M. Schmidt, Loretta Arcangeli, John Ransom, Joshua Trueheart, John P. Manfredi, James R. Broach, Andrew J. Murphy

Research output: Contribution to journalArticle

126 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)1334-1337
Number of pages4
JournalNature Biotechnology
Volume16
Issue number13
DOIs
StatePublished - Dec 1 1998

Fingerprint

Formyl Peptide Receptor
Yeast
Peptides
G-Protein-Coupled Receptors
Yeasts
Chemical activation
Histidine
Peptide Library
Neutrophil Activation
Saccharomyces
HEK293 Cells
Proteins
Neutrophils
Plasmids
Pharmacology
Calcium
Kidney
human FPR2 protein

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

Cite this

Klein, C., Paul, J. I., Sauvé, K., Schmidt, M. M., Arcangeli, L., Ransom, J., ... Murphy, A. J. (1998). Identification of surrogate agonists for the human FPRL-1 receptor by autocrine selection in yeast. Nature Biotechnology, 16(13), 1334-1337. https://doi.org/10.1038/4310
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. / Identification of surrogate agonists for the human FPRL-1 receptor by autocrine selection in yeast. In: Nature Biotechnology. 1998 ; Vol. 16, No. 13. pp. 1334-1337.
@article{dd7ff51b471c4f8389113953178073c6,
title = "Identification of surrogate agonists for the human FPRL-1 receptor by autocrine selection in yeast",
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.",
author = "Christine Klein and Paul, {Jeremy I.} and Karen Sauv{\'e} and Schmidt, {Mary M.} and Loretta Arcangeli and John Ransom and Joshua Trueheart and Manfredi, {John P.} and Broach, {James R.} and Murphy, {Andrew J.}",
year = "1998",
month = "12",
day = "1",
doi = "10.1038/4310",
language = "English (US)",
volume = "16",
pages = "1334--1337",
journal = "Nature Biotechnology",
issn = "1087-0156",
publisher = "Nature Publishing Group",
number = "13",

}

Klein, C, Paul, JI, Sauvé, K, Schmidt, MM, Arcangeli, L, Ransom, J, Trueheart, J, Manfredi, JP, Broach, JR & Murphy, AJ 1998, 'Identification of surrogate agonists for the human FPRL-1 receptor by autocrine selection in yeast', Nature Biotechnology, vol. 16, no. 13, pp. 1334-1337. https://doi.org/10.1038/4310

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

In: Nature Biotechnology, Vol. 16, No. 13, 01.12.1998, p. 1334-1337.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Identification of surrogate agonists for the human FPRL-1 receptor by autocrine selection in yeast

AU - Klein, Christine

AU - Paul, Jeremy I.

AU - Sauvé, Karen

AU - Schmidt, Mary M.

AU - Arcangeli, Loretta

AU - Ransom, John

AU - Trueheart, Joshua

AU - Manfredi, John P.

AU - Broach, James R.

AU - Murphy, Andrew J.

PY - 1998/12/1

Y1 - 1998/12/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=18744426569&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=18744426569&partnerID=8YFLogxK

U2 - 10.1038/4310

DO - 10.1038/4310

M3 - Article

C2 - 9853614

AN - SCOPUS:18744426569

VL - 16

SP - 1334

EP - 1337

JO - Nature Biotechnology

JF - Nature Biotechnology

SN - 1087-0156

IS - 13

ER -

Klein C, Paul JI, Sauvé K, Schmidt MM, Arcangeli L, Ransom J et al. Identification of surrogate agonists for the human FPRL-1 receptor by autocrine selection in yeast. Nature Biotechnology. 1998 Dec 1;16(13):1334-1337. https://doi.org/10.1038/4310