TY - JOUR
T1 - Palmitoylation of the V2 Vasopressin Receptor Carboxyl Tail Enhances β-Arrestin Recruitment Leading to Efficient Receptor Endocytosis and ERK1/ 2 Activation
AU - Charest, Pascale G.
AU - Bouvier, Michel
PY - 2003/10/17
Y1 - 2003/10/17
N2 - A large number of G protein-coupled receptors are palmitoylated on cysteine residues located in their carboxyl tail, but the general role of this post-translational modification remains poorly understood. Here we show that preventing palmitoylation of the V2 vasopressin receptor, by site-directed mutagenesis of cysteines 341 and 342, significantly delayed and decreased both agonist-promoted receptor endocytosis and mitogen-activated protein kinase activation. Pharmacological blockade of receptor endocytosis is without effect on the vasopressin-stimulated mitogen-activated protein kinase activity, excluding the possibility that the reduced kinase activation mediated by the palmitoylation-less mutant could result from altered receptor endocytosis. In contrast, two dominant negative mutants of β-arrestin which inhibit receptor endocytosis also attenuated vasopressin-stimulated mitogen-activated protein kinase activity, suggesting that the scaffolding protein, β-arrestin, represents the common link among receptor palmitoylation, endocytosis, and kinase activation. Co-immunoprecipitation and bioluminescence resonance energy transfer experiments confirmed that inhibiting receptor palmitoylation considerably reduced the vasopressin-stimulated recruitment of β-arrestin to the receptor. Interestingly, the changes in β-arrestin recruitment kinetics were similar to those observed for vasopressin-stimulated receptor endocytosis and mitogen-activated protein kinase activation. Taken together the results indicate that palmitoylation enhances the recruitment of β-arrestin to the activated V2 vasopressin receptor thus facilitating processes requiring the scaffolding action of β-arrestin.
AB - A large number of G protein-coupled receptors are palmitoylated on cysteine residues located in their carboxyl tail, but the general role of this post-translational modification remains poorly understood. Here we show that preventing palmitoylation of the V2 vasopressin receptor, by site-directed mutagenesis of cysteines 341 and 342, significantly delayed and decreased both agonist-promoted receptor endocytosis and mitogen-activated protein kinase activation. Pharmacological blockade of receptor endocytosis is without effect on the vasopressin-stimulated mitogen-activated protein kinase activity, excluding the possibility that the reduced kinase activation mediated by the palmitoylation-less mutant could result from altered receptor endocytosis. In contrast, two dominant negative mutants of β-arrestin which inhibit receptor endocytosis also attenuated vasopressin-stimulated mitogen-activated protein kinase activity, suggesting that the scaffolding protein, β-arrestin, represents the common link among receptor palmitoylation, endocytosis, and kinase activation. Co-immunoprecipitation and bioluminescence resonance energy transfer experiments confirmed that inhibiting receptor palmitoylation considerably reduced the vasopressin-stimulated recruitment of β-arrestin to the receptor. Interestingly, the changes in β-arrestin recruitment kinetics were similar to those observed for vasopressin-stimulated receptor endocytosis and mitogen-activated protein kinase activation. Taken together the results indicate that palmitoylation enhances the recruitment of β-arrestin to the activated V2 vasopressin receptor thus facilitating processes requiring the scaffolding action of β-arrestin.
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U2 - 10.1074/jbc.M306589200
DO - 10.1074/jbc.M306589200
M3 - Article
C2 - 12900404
AN - SCOPUS:0142039784
SN - 0021-9258
VL - 278
SP - 41541
EP - 41551
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 42
ER -