Palmitoylation of cysteine 415 of CB1 receptor affects ligand-stimulated internalization and selective interaction with membrane cholesterol and caveolin 1

Sergio Oddi, Tomasz Maciej Stepniewski, Antonio Totaro, Jana Selent, Lucia Scipioni, Beatrice Dufrusine, Filomena Fezza, Enrico Dainese, Mauro Maccarrone

Research output: Contribution to journalArticlepeer-review

Abstract

We previously demonstrated that CB1 receptor is palmitoylated at cysteine 415, and that such a post-translational modification affects its biological activity. To assess the molecular mechanisms responsible for modulation of CB1 receptor function by S-palmitoylation, in this study biochemical and morphological approaches were paralleled with computational analyses. Molecular dynamics simulations suggested that this acyl chain stabilizes helix 8 as well as the interaction of CB1 receptor with membrane cholesterol. In keeping with these in silico data, experimental results showed that the non-palmitoylated CB1 receptor was unable to interact efficaciously with caveolin 1, independently of its activation state. Moreover, in contrast with the wild-type receptor, the lack of S-palmitoylation in the helix 8 made the mutant CB1 receptor completely irresponsive to agonist-induced effects in terms of both lipid raft partitioning and receptor internalization. Overall, our results support the notion that palmitoylation of cysteine 415 modulates the conformational state of helix 8 and influences the interactions of CB1 receptor with cholesterol and caveolin 1, suggesting that the palmitoyl chain may serve as a functional interface for CB1 receptor localization and function.

Original languageEnglish
Pages (from-to)523-532
Number of pages10
JournalBiochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
Volume1862
Issue number5
DOIs
Publication statusPublished - May 1 2017

Keywords

  • Caveolae
  • Homology modeling
  • Lipid rafts
  • Molecular dynamics
  • Receptor acylation
  • Type-1 cannabinoid receptor

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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