Molecular pharmacology
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Molecular pharmacology · Aug 2004
Interleukin-1beta-induced mucin production in human airway epithelium is mediated by cyclooxygenase-2, prostaglandin E2 receptors, and cyclic AMP-protein kinase A signaling.
We reported recently that interleukin (IL)-1beta exposure resulted in a prolonged increase in MUC5AC mucin production in normal, well differentiated, human tracheobronchial epithelial (NHTBE) cell cultures, without significantly increasing MUC5AC mRNA (Am J Physiol 286:L320-L330, 2004). The goal of the present study was to elucidate the signaling pathways involved in IL-1beta-induced MUC5AC production. We found that IL-1beta increased cyclooxygenase-2 (COX-2) mRNA expression and prostaglandin (PG) E(2) production and that the COX-2 inhibitor celecoxib suppressed IL-1beta-induced MUC5AC production. ⋯ However, neither inhibition of epidermal growth factor receptor (EGFR) activation with the tyrosine kinase inhibitor 4-(3-chloroanilino)-6,7-dimethoxyquinazoline HCl (AG-1478) or EGFR blocking antibody nor inhibition of extracellular signal-regulated kinase/P-38 mitogen activated protein kinases with specific inhibitors blocked IL-1beta stimulation of MUC5AC mucin production. We also observed that tumor necrosis factor (TNF)-alpha, platelet activating factor (PAF), and lipopolysaccharide (LPS) induced COX-2 and increased MUC5AC production that was blocked by celecoxib, suggesting a common signaling pathway of inflammatory mediator-induced MUC5AC production in NHTBE cells. We conclude that the induction of MUC5AC by IL-1beta, TNF-alpha, PAF, and LPS involves COX-2- generated PGE(2), activation of EP2 and/or EP4 receptor(s), and cAMP-PKA-mediated signaling.
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Molecular pharmacology · Aug 2004
Identification of a novel site within G protein alpha subunits important for specificity of receptor-G protein interaction.
Several domains of G protein alpha subunits are implicated in the control of receptor-G protein coupling specificity. Among these are the extreme N-and C-termini, the alpha4/beta6-loops, and the loop linking the N-terminal alpha-helix to the beta1-strand of the ras-like domain. In this study, we illustrate that single-point mutations of a highly conserved glycine residue within the linker I region of the Galpha(q) subunit confers upon the mutant Galpha(q) the ability to be activated by Galpha(i)- and Galpha(s) -coupled receptors, as evidenced by guanosine 5'-O-(3-[(35)S]thio)triphosphate binding and inositol phosphate turnover assays. ⋯ It is noteworthy that both mutant and wild-type Galpha(q) proteins are indistinguishable in their ability to reconstitute a functional Gq-PLCbeta-calcium signaling pathway when cotransfected with the Galpha(q)-coupled neurokinin 1 or muscarinic M3 receptor into mouse embryonic fibroblasts derived from Galpha(q/11) knockout mice. On a three-dimensional model of the receptor-G protein complex, the highly conserved linker I region connecting the helical and the GTPase domain of the Galpha protein is inaccessible to the intracellular surface of the receptors. Our data indicate that receptor-G protein coupling specificity is not exclusively governed by direct receptor-G protein interaction and that it even bypasses the requirement of the extreme C terminus of Galpha, a well accepted receptor recognition domain, suggesting a novel allosteric mechanism for G protein-coupled receptor-G protein selectivity.