Tags

Type your tag names separated by a space and hit enter

Palmitoylation of cysteine 415 of CB1 receptor affects ligand-stimulated internalization and selective interaction with membrane cholesterol and caveolin 1.
Biochim Biophys Acta Mol Cell Biol Lipids. 2017 May; 1862(5):523-532.BB

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.

Links

Publisher Full Text (DOI)

Authors+Show Affiliations

Oddi S
Faculty of Veterinary Medicine, University of Teramo, Teramo, Italy; European Center for Brain Research (CERC)/Santa Lucia Foundation IRCCS, Rome, Italy. Electronic address: soddi@unite.it.
Stepniewski TM
Research Programme on Biomedical Informatics (GRIB), Department of Experimental and Health Sciences of Pompeu Fabra University (UPF), Hospital del Mar Medical Research Institute (IMIM), Barcelona, Spain; Faculty of Chemistry, Biological and Chemical Research Centre, University of Warsaw, Warsaw, Poland.
Totaro A
European Center for Brain Research (CERC)/Santa Lucia Foundation IRCCS, Rome, Italy.
Selent J
Research Programme on Biomedical Informatics (GRIB), Department of Experimental and Health Sciences of Pompeu Fabra University (UPF), Hospital del Mar Medical Research Institute (IMIM), Barcelona, Spain.
Scipioni L
European Center for Brain Research (CERC)/Santa Lucia Foundation IRCCS, Rome, Italy; Department of Medicine, Campus Bio-Medico University of Rome, Rome, Italy.
Dufrusine B
Faculty of Bioscience and Technology for Food Agriculture and Environment, University of Teramo, Teramo, Italy.
Fezza F
European Center for Brain Research (CERC)/Santa Lucia Foundation IRCCS, Rome, Italy; Department of Experimental Medicine and Surgery, Tor Vergata University of Rome, Rome, Italy.
Dainese E
European Center for Brain Research (CERC)/Santa Lucia Foundation IRCCS, Rome, Italy; Faculty of Bioscience and Technology for Food Agriculture and Environment, University of Teramo, Teramo, Italy.
Maccarrone M
European Center for Brain Research (CERC)/Santa Lucia Foundation IRCCS, Rome, Italy; Department of Medicine, Campus Bio-Medico University of Rome, Rome, Italy. Electronic address: m.maccarrone@unicampus.it.

MeSH

Caveolin 1Cell LineCholesterolCysteineHEK293 CellsHumansLigandsLipoylationMembrane MicrodomainsMolecular Dynamics SimulationMutationPalmitic AcidProtein BindingProtein ConformationProtein Interaction MapsReceptor, Cannabinoid, CB1

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

28215712

Citation

Oddi, Sergio, et al. "Palmitoylation of Cysteine 415 of CB1 Receptor Affects Ligand-stimulated Internalization and Selective Interaction With Membrane Cholesterol and Caveolin 1." Biochimica Et Biophysica Acta. Molecular and Cell Biology of Lipids, vol. 1862, no. 5, 2017, pp. 523-532.
Oddi S, Stepniewski TM, Totaro A, et al. Palmitoylation of cysteine 415 of CB1 receptor affects ligand-stimulated internalization and selective interaction with membrane cholesterol and caveolin 1. Biochim Biophys Acta Mol Cell Biol Lipids. 2017;1862(5):523-532.
Oddi, S., Stepniewski, T. M., Totaro, A., Selent, J., Scipioni, L., Dufrusine, B., Fezza, F., Dainese, E., & Maccarrone, M. (2017). Palmitoylation of cysteine 415 of CB1 receptor affects ligand-stimulated internalization and selective interaction with membrane cholesterol and caveolin 1. Biochimica Et Biophysica Acta. Molecular and Cell Biology of Lipids, 1862(5), 523-532. https://doi.org/10.1016/j.bbalip.2017.02.004
Oddi S, et al. Palmitoylation of Cysteine 415 of CB1 Receptor Affects Ligand-stimulated Internalization and Selective Interaction With Membrane Cholesterol and Caveolin 1. Biochim Biophys Acta Mol Cell Biol Lipids. 2017;1862(5):523-532. PubMed PMID: 28215712.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Palmitoylation of cysteine 415 of CB1 receptor affects ligand-stimulated internalization and selective interaction with membrane cholesterol and caveolin 1. AU - Oddi,Sergio, AU - Stepniewski,Tomasz Maciej, AU - Totaro,Antonio, AU - Selent,Jana, AU - Scipioni,Lucia, AU - Dufrusine,Beatrice, AU - Fezza,Filomena, AU - Dainese,Enrico, AU - Maccarrone,Mauro, Y1 - 2017/02/12/ PY - 2016/09/28/received PY - 2017/02/02/revised PY - 2017/02/10/accepted PY - 2017/2/22/pubmed PY - 2017/7/14/medline PY - 2017/2/21/entrez KW - Caveolae KW - Homology modeling KW - Lipid rafts KW - Molecular dynamics KW - Receptor acylation KW - Type-1 cannabinoid receptor SP - 523 EP - 532 JF - Biochimica et biophysica acta. Molecular and cell biology of lipids JO - Biochim Biophys Acta Mol Cell Biol Lipids VL - 1862 IS - 5 N2 - 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. SN - 1388-1981 UR - https://www.unboundmedicine.com/medline/citation/28215712/Palmitoylation_of_cysteine_415_of_CB1_receptor_affects_ligand_stimulated_internalization_and_selective_interaction_with_membrane_cholesterol_and_caveolin_1_ DB - PRIME DP - Unbound Medicine ER -