Proteinase-activated receptor-2 mediated inhibition of TNFα-stimulated JNK activation — A novel paradigm for Gq/11 linked GPCRs

McIntosh, K., Cunningham, M.R., Cadalbert, L., Lockhart, J., Boyd, G., Ferrell, W.R. and Plevin, R. (2010) Proteinase-activated receptor-2 mediated inhibition of TNFα-stimulated JNK activation — A novel paradigm for Gq/11 linked GPCRs. Cellular Signalling, 22(2), pp. 265-273. (doi: 10.1016/j.cellsig.2009.09.028)

Abstract

In this study we examined the potential for PAR2 and TNF[alpha] to synergise at the level of MAP kinase signalling in PAR2 expressing NCTC2544 cells. However, to our surprise we found that activation of PAR2 by trypsin or the specific activating peptide SLIGKV-OH strongly inhibited both the phosphorylation and activity of JNK. In contrast neither p38 MAP kinase nor ERK activation was affected although TNF[alpha] stimulated I[kappa]B[alpha] loss was partially reversed. The inhibitory effect was not observed in parental cells nor in cells expressing PAR4, however inhibition was reversed by pre-incubation with the novel PAR2 antagonist K14585, suggesting that the effect is specific for PAR2 activation. SLIGKV-OH was found to be more potent in inhibiting TNF[alpha]-induced JNK activation than in stimulating JNK alone, suggesting agonist-directed signalling. The PKC activator PMA, also mimicked the inhibitory effect of SLIGKV-OH, the effects of both agents were reversed by pre-treatment with the PKC inhibitor, GF109203X. Furthermore, incubation with the novel Gq/11 inhibitor YM25480 also reversed PAR2 mediated inhibition. Activation of PAR2 was found to disrupt TNFR1 binding to RIP and TRADD and this was reversed by both GF109203X and YM25480. A similar mode of inhibition observed in HUVECs through PAR2 or P2Y2 receptors demonstrates the potential of a novel paradigm for GPCRs linked to Gq/11, in mediating inhibition of TNF[alpha]-stimulated JNK activation. This has important implications in assessing the role of GPCRs in inflammation and other conditions

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Ferrell, Professor William
• Authors: McIntosh, K., Cunningham, M.R., Cadalbert, L., Lockhart, J., Boyd, G., Ferrell, W.R., and Plevin, R.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Journal Name: Cellular Signalling
• ISSN: 0898-6568