Activation of protease-activated receptor 2 (PAR2) and release intracellular calcium by mast cell tryptase
Activation of protease-activated receptor 2 (PAR2) and release intracellular calcium by mast cell tryptase
Introduction: Mast cell tryptase can be released in substantial quantities at sites of allergic inflammation. Pro-inflammatory actions mediated through PAR2 have been proposed, but subcellular mechanisms of receptor activation still unclear. Our aim is to investigate the intracellular mechanisms following tryptase activation of PAR2.
METHODS: Kirsten Murine Sarcoma Virus transformed rat kidney epithelial cells transfected with human PAR2 (KNRKt-PAR2) were employed. Expression of PAR2 was investigated using immunoflorescent staining. Following addition of tryptase or other agents, calcium flux was measured using a fluorescence based micro-plate procedure.
RESULTS: Trypsin and the PAR2 peptide agonist (SLIGKV-NH2) were markedly able to stimulate release of intracellular calcium in the KNRKt cells. Addition of tryptase was associated with a concentration-dependent increase in the intracellular calcium. Pre-treatment of the cells with pertussis toxin, G protein-coupled receptors inhibitor was abolished the effect of all agents under investigation on calcium mobilization. Incubation of cells with trypsin was found to prevent the effect of tryptase and vice versa. PAR2 receptors Response to the peptide agonist was greatly reduced following trypsin treatment as compared to tryptase.
CONCLUSIONS: Mechanism of tryptase activation to PAR2 may involve mobilization of calcium inside the target cells, the effect that mimics that of trypsin but with less potency.
Khedr, M.E.M.S.
37c876ff-7226-4d51-bd20-dfe0bbca3d73
9 November 2016
Khedr, M.E.M.S.
37c876ff-7226-4d51-bd20-dfe0bbca3d73
Khedr, M.E.M.S.
(2016)
Activation of protease-activated receptor 2 (PAR2) and release intracellular calcium by mast cell tryptase.
Daniel Turnberg Travel Fellowship Scheme Alumni Conference, , Nicosia, Cyprus.
09 - 11 Nov 2016.
Record type:
Conference or Workshop Item
(Poster)
Abstract
Introduction: Mast cell tryptase can be released in substantial quantities at sites of allergic inflammation. Pro-inflammatory actions mediated through PAR2 have been proposed, but subcellular mechanisms of receptor activation still unclear. Our aim is to investigate the intracellular mechanisms following tryptase activation of PAR2.
METHODS: Kirsten Murine Sarcoma Virus transformed rat kidney epithelial cells transfected with human PAR2 (KNRKt-PAR2) were employed. Expression of PAR2 was investigated using immunoflorescent staining. Following addition of tryptase or other agents, calcium flux was measured using a fluorescence based micro-plate procedure.
RESULTS: Trypsin and the PAR2 peptide agonist (SLIGKV-NH2) were markedly able to stimulate release of intracellular calcium in the KNRKt cells. Addition of tryptase was associated with a concentration-dependent increase in the intracellular calcium. Pre-treatment of the cells with pertussis toxin, G protein-coupled receptors inhibitor was abolished the effect of all agents under investigation on calcium mobilization. Incubation of cells with trypsin was found to prevent the effect of tryptase and vice versa. PAR2 receptors Response to the peptide agonist was greatly reduced following trypsin treatment as compared to tryptase.
CONCLUSIONS: Mechanism of tryptase activation to PAR2 may involve mobilization of calcium inside the target cells, the effect that mimics that of trypsin but with less potency.
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Published date: 9 November 2016
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Daniel Turnberg Travel Fellowship Scheme Alumni Conference, , Nicosia, Cyprus, 2016-11-09 - 2016-11-11
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Local EPrints ID: 432924
URI: http://eprints.soton.ac.uk/id/eprint/432924
PURE UUID: c3d42b75-6462-4b2b-b5b1-6b474d8c6838
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Date deposited: 01 Aug 2019 16:30
Last modified: 13 Dec 2021 03:09
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Author:
M.E.M.S. Khedr
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