University of Hull logo

Palmitoylation of human proteinase-activated receptor-2 differentially regulates receptor-triggered ERK1/2 activation, calcium signalling and endocytosis

Xiao, Yu Pei; Morice, Alyn H.; Sadofsky, Laura R.; Botham, Andrew; Guo, Xiaodan; Xiao, Yu Pei; Morice, Alyn H.; Compton, Steven J.; Compton, Steven J.; Sadofsky, Laura R.

Authors

Yu Pei Xiao

Alyn H. Morice

Laura R. Sadofsky

Andrew Botham

Xiaodan Guo

Yu Pei Xiao

Alyn H. Morice

Steven J. Compton

Steven J. Compton

Dr Laura Sadofsky L.R.Sadofsky@hull.ac.uk
Lecturer in Respiratory Medicine/ Academic lead for postgraduate training in HYMS

Abstract

hPAR2 (human proteinase-activated receptor-2) is a member of the novel family of proteolytically activated GPCRs (G-protein-coupled receptors) termed PARs (proteinase-activated receptors). Previous pharmacological studies have found that activation of hPAR2 by mast cell tryptase can be regulated by receptor N-terminal glycosylation. In order to elucidate other post-translational modifications of hPAR2 that can regulate function, we have explored the functional role of the intracellular cysteine residue Cys361. We have demonstrated, using autoradiography, that Cys361 is the primary palmitoylation site of hPAR2. The hPAR2C361A mutant cell line displayed greater cell-surface expression compared with the wt (wild-type)-hPAR2-expressing cell line. hPAR2C361A also showed a decreased sensitivity and efficacy (intracellular calcium signalling) towards both trypsin and SLIGKV. In stark contrast, hPAR2C361A triggered greater and more prolonged ERK (extracellular-signal-regulated kinase) phosphorylation compared with that of wt-hPAR2 possibly through Gi, since pertussis toxin inhibited the ability of this receptor to activate ERK. Finally, flow cytometry was utilized to assess the rate and extent of receptor internalization following agonist challenge. hPAR2C361A displayed faster internalization kinetics following trypsin activation compared with wt-hPAR2, whereas SLIGKV had a negligible effect on internalization for either receptor. In conclusion, palmitoylation plays an important role in the regulation of PAR2 expression, agonist sensitivity, desensitization and internalization.

Journal Article Type Article
Publication Date Sep 1, 2011
Journal Biochemical journal
Print ISSN 1470-8728
Electronic ISSN 1470-8728
Publisher Portland Press
Peer Reviewed Peer Reviewed
Volume 438
Issue 2
Pages 359-367
Institution Citation Botham, A., Guo, X., Xiao, Y. P., Morice, A. H., Compton, S., & Sadofsky, L. R. (2011). Palmitoylation of human proteinase-activated receptor-2 differentially regulates receptor-triggered ERK1/2 activation, calcium signalling and endocytosis. Biochemical Journal, 438(2), (359-367). doi:10.1042/BJ20101958. ISSN 1470-8728
DOI https://doi.org/10.1042/BJ20101958
Keywords Cell Biology; Biochemistry; Molecular Biology
Publisher URL http://www.biochemj.org/bj/438/bj4380359.htm
Additional Information Copy of article first published in Biochemical journal, 2011, v.438, issue 2

Files



You might also like


Downloadable Citations