Proteinase-activated receptor-2 mediated inhibition of TNF alpha-stimulated JNK activation: A novel paradigm for G(q/11) linked GPCRs

Kathryn McIntosh, Margaret R. Cunningham, Laurence Cadalbert, John Lockhart, Gary Boyd, William R. Ferrell, Robin Plevin

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7 Citations (Scopus)

Abstract

In this study we examined the potential for PAR(2) and TNF alpha to synergise at the level of MAP kinase signalling in PAR(2) expressing NCTC2544 cells. However, to our surprise we found that activation of PAR(2) 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 PAR(4), however inhibition was reversed by pre-incubation with the novel PAR(2) antagonist K14585. suggesting that the effect is specific for PAR(2) activation. SLIGV-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, and the effects of both agents were reversed by pretreatment with the PKC inhibitor, GF109203X. Furthermore, incubation with the novel G(q/11) inhibitor YM25480 also reversed PAR(2) mediated inhibition. Activation of PAR(2) 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 PAR(2) or P2Y2 receptors demonstrates the potential of a novel paradigm for GPCRs linked to G(q/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.
Original languageEnglish
Pages (from-to)265-273
JournalCellular Signalling
Volume22
Issue number2
DOIs
Publication statusPublished - Feb 2010

Keywords

  • Proteinase-activated receptor 2
  • c-jun N-terminal protein kinase
  • TNF alpha
  • Protein kinase C

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