PDZ-dependent Activation of Nitric-oxide Synthases by the Serotonin 2B Receptor

• Published in: The Journal of biological chemistry Vol. 275; no. 13; pp. 9324 - 9331
• Main Authors: Manivet, Philippe, Mouillet-Richard, Sophie, Callebert, Jacques, Nebigil, Canan G., Maroteaux, Luc, Hosoda, Syun, Kellermann, Odile, Launay, Jean-Marie
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 31-03-2000; American Society for Biochemistry and Molecular Biology
• Subjects: Amino Acid Sequence; Base Sequence; Cyclic GMP - metabolism; DNA Primers; Enzyme Activation; Isoenzymes - metabolism; Mastomys natalensis; Molecular Sequence Data; Nitric Oxide - biosynthesis; Nitric Oxide Synthase - metabolism; Protein Binding; Receptor, Serotonin, 5-HT2B; Receptors, Serotonin - physiology; rodents
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.275.13.9324

Taking advantage of three cellular systems, we established that 5-HT2B receptors are coupled with NO signaling pathways. In the 1C11 serotonergic cell line andMastomys natalensis carcinoid cells, which naturally express the 5-HT2B receptor, as well as in transfected LMTK− fibroblasts, stimulation of the 5-HT2Breceptor triggers intracellular cGMP production through dual activation of constitutive nitric-oxide synthase (cNOS) and inducible NOS (iNOS). The group I PDZ motif at the C terminus of the 5-HT2Breceptor is required for recruitment of the cNOS and iNOS transduction pathways. Indeed, the 5-HT2B receptor-mediated NO coupling is abolished not only upon introduction of a competitor C-terminal 5-HT2B peptide in the three cell types but also in LMTK− fibroblasts expressing a receptor C-terminally truncated or harboring a point mutation within the PDZ domain. The occurrence of a direct functional coupling between the receptor and cNOS activity is supported by highly significant correlations between the binding constants of drugs on the receptor and their effects on cNOS activity. The 5-HT2B/iNOS coupling mechanisms appear more complex because neutralization of endogenous Gα13 by specific antibodies cancels the cellular iNOS response while not interfering with cNOS activities. These findings may shed light on physiological links between the 5-HT2B receptor and NO and constitute the first demonstration that PDZ interactions participate in downstream transductional pathways of a G protein-coupled receptor.