期刊
JOURNAL OF PHARMACOLOGICAL AND TOXICOLOGICAL METHODS
卷 88, 期 -, 页码 72-78出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.vascn.2017.07.003
关键词
Dynamic mass redistribution; GPCR; FPR1; FPR2; Label-free assay; Neutrophil; Potency; Signalling
资金
- Carlsberg Foundation
- European Research Council [DE-ORPHAN 639125]
- Lundbeck Foundation [R169-2013-16327]
- Lundbeck Foundation
- Lundbeck Foundation [R163-2013-16327] Funding Source: researchfish
Introduction: The label-free dynamic mass redistribution-based assay (DMR) is a powerful method for studying signalling pathways of G protein-coupled receptors (GPCRs). Herein we present the label-free DMR assay as a robust readout for pharmacological characterization of formyl peptide receptors (FPRs) in human neutrophils. Methods: Neutrophils were isolated from fresh human blood and their responses to FPR1 and FPR2 agonists, i.e. compound 43, fMLF and WKYMVm were measured in a label-free DMR assay using Epic Benchtop System from Corning (R). Obtained DMR traces were used to calculate agonist potencies. Results: The potencies (pEC(50)) of fMLF, WKYMVm and compound 43, determined on human neutrophils using the label-free DMR assay were 8.63, 7.76 and 5.92, respectively. The DMR response to fMLF, but not WKYMVm and compound 43 could be blocked by the FPR1-specific antagonist cyclosporin H. Discussion: We conclude that the DMR assay can be used, and complements more traditional methods, to study the signalling and pharmacology of endogenous FPR receptors in human neutrophils.
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