Mapping transmembrane residues of proteinase activated recpetor 2 (PAR2) that influence ligand-modulated calcium signaling

Suen, J.Y., Adams, M. N., Lim, J., Madala, P.K., Xu, W, Cotterell, A., He, Y., Yua, Mei-Kwan, Hooper, J. D. and Fairlie, D.P. (2017) Mapping transmembrane residues of proteinase activated recpetor 2 (PAR2) that influence ligand-modulated calcium signaling. Pharmacological Research, 117 328-342. doi:10.1016/j.phrs.2016.12.020

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Author Suen, J.Y.
Adams, M. N.
Lim, J.
Madala, P.K.
Xu, W
Cotterell, A.
He, Y.
Yua, Mei-Kwan
Hooper, J. D.
Fairlie, D.P.
Title Mapping transmembrane residues of proteinase activated recpetor 2 (PAR2) that influence ligand-modulated calcium signaling
Formatted title
Mapping transmembrane residues of proteinase activated recpetor 2 (PAR2) that influence ligand-modulated calcium signaling
Journal name Pharmacological Research   Check publisher's open access policy
ISSN 1043-6618
1096-1186
Publication date 2017-03-01
Year available 2016
Sub-type Article (original research)
DOI 10.1016/j.phrs.2016.12.020
Open Access Status File (Author Post-print)
Volume 117
Start page 328
End page 342
Total pages 52
Place of publication London, United Kingdom
Publisher Academic Press
Language eng
Formatted abstract
Proteinase-activated receptor 2 (PAR2) is a G protein-coupled receptor involved in metabolism, inflammation, and cancers. It is activated by proteolysis, which exposes a nascent N-terminal sequence that becomes a tethered agonist. Short synthetic peptides corresponding to this sequence also activate PAR2, while small organic molecules show promising PAR2 antagonism. Developing PAR2 ligands into pharmaceuticals is hindered by a lack of knowledge of how synthetic ligands interact with and differentially modulate PAR2. Guided by PAR2 homology modeling and ligand docking based on bovine rhodopsin, followed by cross-checking with newer PAR2 models based on ORL-1 and PAR1, site-directed mutagenesis of PAR2 was used to investigate the pharmacology of three agonists (two synthetic agonists and trypsin-exposed tethered ligand) and one antagonist for modulation of PAR2 signaling. Effects of 28 PAR2 mutations were examined for PAR2-mediated calcium mobilization and key mutants were selected for measuring ligand binding. Nineteen of twenty-eight PAR2 mutations reduced the potency of at least one ligand by >10-fold. Key residues mapped predominantly to a cluster in the transmembrane (TM) domains of PAR2, differentially influence intracellular Ca2+ induced by synthetic agonists versus a native agonist, and highlight subtly different TM residues involved in receptor activation. This is the first evidence highlighting the importance of the PAR2 TM regions for receptor activation by synthetic PAR2 agonists and antagonists. The trypsin-cleaved N-terminus that activates PAR2 was unaffected by the same residues as synthetic peptides, challenging the widespread practice of substituting peptides for proteases to characterize PAR2 physiology.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

 
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Created: Thu, 05 Jan 2017, 02:04:01 EST by Susan Allen on behalf of Institute for Molecular Bioscience