Abstract
In order to properly interpret receptor inhibition experiments, the precise receptor specificities of the employed antagonists are of crucial importance. Lately, a great number of agonists for various formyl peptide receptors have been identified using a selection of antagonists. However, some confusion exists as to the precise receptor specificities of many of these antagonists. We have investigated the effects of formyl peptide receptor family antagonists on the neutrophil response induced by agonists for the formyl peptide receptor (FPR) and the formyl peptide receptor like 1 (FPRL1). To determine FPR- and FPRL1-specific interactions, these antagonists should not be used at used at concentrations above 10 μM. Signaling through FPR was inhibited by low concentrations of the antagonists cyclosporin H, Boc-MLF (also termed Boc-1), and Boc-FLFLFL (also termed Boc-2), while higher concentrations also partly inhibited the signaling through FPRL1. The antagonist WRWWWW (WRW4) specifically inhibited the signaling through FPRL1 at low concentrations but at high concentrations also partly the signaling through FPR. Based on the difference in potency of cyclosporin H and the two Boc-peptides, we suggest using cyclosporin H as a specific inhibitor for FPR. To specifically inhibit the FPRL1 response the antagonist WRW4 should be used.
Similar content being viewed by others
References
Schiffmann, E., B. A. Corcoran, and S. M. Wahl. 1975. N-formylmethionyl peptides as chemoattractants for leucocytes. Proc. Natl. Acad. Sci. USA 72:1059–1062.
Miller, A. F., and J. J. Falke. 2004. Chemotaxis receptors and signaling. Adv. Protein Chem. 68:393–444.
Ye, R. D., and F. Boulay. 1997. Structure and function of leukocyte chemoattractant receptors. Adv. Pharmacol. 39:221–289.
Ronald J U., R.S. 1999. Chemoattractant stimulus–response coupling. In: Inflammation: Basic Priciples and Clinical Correlates, R. S. John I G., ed. Lippincott Williams & Wilkins, Philadelphia 607–626.
Le, Y., P. M. Murphy, and J. M. Wang. 2002. Formyl-peptide receptors revisited. Trends. Immunol. 23:541–548.
Boulay, F., M. Tardif, L. Brouchon, and P. Vignais. 1990. Synthesis and use of a novel N-formyl peptide derivative to isolate a human N-formyl peptide receptor cDNA. Biochem. Biophys. Res. Commun. 168:1103–1109.
Ye, R. D., S. L. Cavanagh, O. Quehenberger, E. R. Prossnitz, and C. G. Cochrane. 1992. Isolation of a cDNA that encodes a novel granulocyte N-formyl peptide receptor. Biochem. Biophys. Res. Commun. 184:582–589.
Fu, H., C. Dahlgren, and J. Bylund. 2003. Subinhibitory concentrations of the deformylase inhibitor actinonin increase bacterial release of neutrophil-activating peptides: A new approach to antimicrobial chemotherapy. Antimicrob. Agents Chemother. 47:2545–2550.
Rabiet, M. J., E. Huet, and F. Boulay. 2005. Human mitochondria-derived N-formylated peptides are novel agonists equally active on FPR and FPRL1, while Listeria monocytogenes-derived peptides preferentially activate FPR. Eur. J. Immunol. 35:2486–2495.
Freer, R. J., A. R. Day, J. A. Radding, E. Schiffmann, S. Aswanikumar, H. J. Showell, and E. L. Becker. 1980. Further studies on the structural requirements for synthetic peptide chemoattractants. Biochemistry. 19:2404–2410.
Betten, A., C. Dahlgren, S. Hermodsson, and K. Hellstrand. 2003. Histamine inhibits neutrophil NADPH oxidase activity triggered by the lipoxin A4 receptor-specific peptide agonist Trp-Lys-Tyr-Met-Val-Met. Scand. J. Immunol. 58:321–326.
Fu, H., L. Bjorkman, P. Janmey, A. Karlsson, J. Karlsson, C. Movitz, and C. Dahlgren. 2004. The two neutrophil members of the formylpeptide receptor family activate the NADPH-oxidase through signals that differ in sensitivity to a gelsolin derived phosphoinositide-binding peptide. BMC Cell Biol. 5:50.
Fu, H., J. Karlsson, J. Bylund, C. Movitz, A. Karlsson, and C. Dahlgren. 2006. Ligand recognition and activation of formyl peptide receptors in neutrophils. J. Leukoc. Biol. 79:247–256.
Migeotte, I., D. Communi, and M. Parmentier. 2006. Formyl peptide receptors: A promiscuous subfamily of G protein-coupled receptors controlling immune responses. Cytokine Growth Factor Rev. 17:501–519.
Perretti, M., N. Chiang, M. La, I. M. Fierro, S. Marullo, S. J. Getting, E. Solito, and C. N. Serhan. 2002. Endogenous lipid- and peptide-derived anti-inflammatory pathways generated with glucocorticoid and aspirin treatment activate the lipoxin A4 receptor. Nat. Med. 8:1296–1302.
Babbin, B. A., W. Y. Lee, C. A. Parkos, L. M. Winfree, A. Akyildiz, M. Perretti, and A. Nusrat. 2006. Annexin I regulates SKCO-15 cell invasion by signaling through formyl peptide receptors. J. Biol. Chem. 281:19588–19599.
Hayhoe, R. P., A. M. Kamal, E. Solito, R. J. Flower, D. Cooper, and M. Perretti. 2006. Annexin 1 and its bioactive peptide inhibit neutrophil–endothelium interactions under flow: Indication of distinct receptor involvement. Blood. 107:2123–2130.
Machado, F. S., J. E. Johndrow, L. Esper, A. Dias, A. Bafica, C. N. Serhan, and J. Aliberti. 2006. Anti-inflammatory actions of lipoxin A4 and aspirin-triggered lipoxin are SOCS-2 dependent. Nat. Med. 12:330–334.
Gavins, F. N., J. Dalli, R. J. Flower, D. N. Granger, and M. Perretti. 2007. Activation of the annexin 1 counter-regulatory circuit affords protection in the mouse brain microcirculation. Faseb J. 21:1751–1758 (Abstract)
John, C. D., V. Sahni, D. Mehet, J. F. Morris, H. C. Christian, M. Perretti, R. J. Flower, E. Solito and J. C. Buckingham. 2007. Formyl peptide receptors and the regulation of ACTH secretion: Targets for annexin A1, lipoxins, and bacterial peptides. Faseb J. 21:1037–1046 (Abstract).
Boyum, A., D. Lovhaug, L. Tresland and E. M. Nordlie. 1991. Separation of leucocytes: Improved cell purity by fine adjustments of gradient medium density and osmolality. Scand. J. Immunol. 34:697–712.
Dahlgren, C.,and A. Karlsson. 1999. Respiratory burst in human neutrophils. J. Immunol. Methods. 232:3–14.
Lundqvist, H.and C. Dahlgren. 1996. Isoluminol-enhanced chemiluminescence: A sensitive method to study the release of superoxide anion from human neutrophils. Free Radic. Biol. Med. 20:785–792.
Wenzel-Seifert, K., and R. Seifert. 1993. Cyclosporin H is a potent and selective formyl peptide receptor antagonist. Comparison with N-t-butoxycarbonyl-l-phenylalanyl-l-leucyl-l-phenylalanyl-l-leucyl-l-phenylalanine and cyclosporins A, B, C, D, and E. J. Immunol. 150:4591–4599.
Seifert, R., and K. Wenzel-Seifert. 2003. The human formyl peptide receptor as model system for constitutively active G-protein-coupled receptors. Life Sci. 73:2263–2280.
Christophe, T., A. Karlsson, C. Dugave, M. J. Rabiet, F. Boulay, and C. Dahlgren. 2001. The synthetic peptide Trp-Lys-Tyr-Met-Val-Met-NH2 specifically activates neutrophils through FPRL1/lipoxin A4 receptors and is an agonist for the orphan monocyte-expressed chemoattractant receptor FPRL2. J. Biol. Chem. 276:21585–21593.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Stenfeldt, AL., Karlsson, J., Wennerås, C. et al. Cyclosporin H, Boc-MLF and Boc-FLFLF are Antagonists that Preferentially Inhibit Activity Triggered Through the Formyl Peptide Receptor. Inflammation 30, 224–229 (2007). https://doi.org/10.1007/s10753-007-9040-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10753-007-9040-4