Palmitoylation of the Vasopressin V1a Receptor Reveals Different Conformational Requirements for Signaling, Agonist-induced Receptor Phosphorylation, and Sequestration

Stuart R. Hawtin, Andrew B. Tobin, Smita Patel, Mark Wheatley

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Abstract

In this study, we establish that the V1a vasopressin receptor (V1aR) is palmitoylated, and we show that this modification has an important functional role. Palmitoylation of the V1aR occurs within the Cys371/Cys372 couplet located in the proximal C-terminal tail domain. Substitution of these residues in a [C371G/C372G]V 1aR construct effectively disrupted receptor palmitoylation. Our data also indicate an additional palmitoylation site at another locus in the receptor, as yet undefined. [3H]Palmitate incorporation was agonist-sensitive and increased following exposure to [Arg 8]vasopressin (AVP). Given the hydrophobic nature of the acyl chain, palmitoylation of the C terminus of G-protein-coupled receptors has been proposed to form an additional intracellular loop. Consequently, palmitoylation/depalmitoylation will have a profound effect on the local conformation of this domain. The V1aR palmitoylation status regulated both phosphorylation and sequestration of the receptor, and furthermore, palmitoylation, phosphorylation, and sequestration were all regulated by AVP. The palmitoylation-defective construct [C371G/C372G]V 1aR exhibited decreased phosphorylation compared to wild-type V 1aR, under both basal and AVP-stimulated conditions, and was sequestered at a faster rate. In contrast, the binding of four different classes of ligand and intracellular signaling were not affected by palmitoylation. This study therefore establishes that there are different conformational requirements for signaling, agonist-induced phosphorylation, and sequestration of the V1aR.

Original languageEnglish
Pages (from-to)38139-38146
Number of pages8
JournalJournal of Biological Chemistry
Volume276
Issue number41
Early online date5 Oct 2001
Publication statusPublished - 12 Oct 2001
Externally publishedYes

Bibliographical note

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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