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Downregulation of 5-HT7 serotonin receptors by the atypical antipsychotics clozapine and olanzapine: role of motifs in the C-terminal domain and interaction with GASP-1

(2015) ACS CHEMICAL NEUROSCIENCE. 6(7). p.1206-1218
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Abstract
The human 5-HT7 serotonin receptor, a G-protein-coupled receptor (GPCR), activates adenylyl cyclase constitutively and upon agonist activation. Biased ligands differentially activate 5-HT7 serotonin receptor desensitization, internalization and degradation in addition to G protein activation. We have previously found that the atypical antipsychotics clozapine and olanzapine inhibited G protein Lysosome activation and, surprisingly, induced both internalization and lysosomal degradation of 5-HT7 receptors. Here, we aimed to determine the mechanism of clozapine- and olanzapine-mediated degradation of 5-HT7 receptors. In the C-terminus of the 5-HT7 receptor, we identified two YXX Phi motifs, LR residues, and a palmitoylated cysteine anchor as potential sites involved in receptor trafficking to lysosomes followed by receptor degradation. Mutating either of these sites inhibited clozapine- and olanzapine-mediated degradation of 5-HT7 receptors and also interfered with G protein activation. In addition, we tested whether receptor degradation was mediated by the GPCR-associated sorting protein-1 (GASP-1). We show that GASP-1 binds the 5-HT7 receptor and regulates the clozapine-mediated degradation. Mutations of the identified motifs and residues, located in or close to Helix-VIII of the 5-HT7 receptor, modified antipsychotic-stimulated binding of proteins (such as GASP-1), possibly by altering the flexibility of Helix-VIII, and also interfered with G protein activation. Taken together, our data demonstrate that binding of clozapine or olanzapine to the 5-HT7 receptor leads to antagonist-mediated lysosomal degradation by exposing key residues in the C-terminal tail that interact with GASP-1.
Keywords
clozapine, 5-HT7 receptor, olanzapine, GASP-1, lysosome, Helix-VIII, PROTEIN-COUPLED RECEPTORS, 4TH CYTOPLASMIC LOOP, FUNCTIONAL SELECTIVITY, SORTING PROTEINS, INTRACELLULAR TRAFFICKING, ADENYLYL-CYCLASE, SPLICE VARIANTS, DRUG DISCOVERY, D2 RECEPTORS, EFFICACY

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Chicago
Manfra, Omella, Kathleen Van Craenenbroeck, Kamila Skieterska, Thomas Frimurer, Thue W Schwartz, Finn Olav Levy, and Kjetil Wessel Andressen. 2015. “Downregulation of 5-HT7 Serotonin Receptors by the Atypical Antipsychotics Clozapine and Olanzapine: Role of Motifs in the C-terminal Domain and Interaction with GASP-1.” Acs Chemical Neuroscience 6 (7): 1206–1218.
APA
Manfra, O., Van Craenenbroeck, K., Skieterska, K., Frimurer, T., Schwartz, T. W., Levy, F. O., & Andressen, K. W. (2015). Downregulation of 5-HT7 serotonin receptors by the atypical antipsychotics clozapine and olanzapine: role of motifs in the C-terminal domain and interaction with GASP-1. ACS CHEMICAL NEUROSCIENCE, 6(7), 1206–1218.
Vancouver
1.
Manfra O, Van Craenenbroeck K, Skieterska K, Frimurer T, Schwartz TW, Levy FO, et al. Downregulation of 5-HT7 serotonin receptors by the atypical antipsychotics clozapine and olanzapine: role of motifs in the C-terminal domain and interaction with GASP-1. ACS CHEMICAL NEUROSCIENCE. 2015;6(7):1206–18.
MLA
Manfra, Omella et al. “Downregulation of 5-HT7 Serotonin Receptors by the Atypical Antipsychotics Clozapine and Olanzapine: Role of Motifs in the C-terminal Domain and Interaction with GASP-1.” ACS CHEMICAL NEUROSCIENCE 6.7 (2015): 1206–1218. Print.
@article{7215220,
  abstract     = {The human 5-HT7 serotonin receptor, a G-protein-coupled receptor (GPCR), activates adenylyl cyclase constitutively and upon agonist activation. Biased ligands differentially activate 5-HT7 serotonin receptor desensitization, internalization and degradation in addition to G protein activation. We have previously found that the atypical antipsychotics clozapine and olanzapine inhibited G protein Lysosome activation and, surprisingly, induced both internalization and lysosomal degradation of 5-HT7 receptors. Here, we aimed to determine the mechanism of clozapine- and olanzapine-mediated degradation of 5-HT7 receptors. In the C-terminus of the 5-HT7 receptor, we identified two YXX Phi motifs, LR residues, and a palmitoylated cysteine anchor as potential sites involved in receptor trafficking to lysosomes followed by receptor degradation. Mutating either of these sites inhibited clozapine- and olanzapine-mediated degradation of 5-HT7 receptors and also interfered with G protein activation. In addition, we tested whether receptor degradation was mediated by the GPCR-associated sorting protein-1 (GASP-1). We show that GASP-1 binds the 5-HT7 receptor and regulates the clozapine-mediated degradation. Mutations of the identified motifs and residues, located in or close to Helix-VIII of the 5-HT7 receptor, modified antipsychotic-stimulated binding of proteins (such as GASP-1), possibly by altering the flexibility of Helix-VIII, and also interfered with G protein activation. Taken together, our data demonstrate that binding of clozapine or olanzapine to the 5-HT7 receptor leads to antagonist-mediated lysosomal degradation by exposing key residues in the C-terminal tail that interact with GASP-1.},
  author       = {Manfra, Omella and Van Craenenbroeck, Kathleen and Skieterska, Kamila and Frimurer, Thomas and Schwartz, Thue W and Levy, Finn Olav and Andressen, Kjetil Wessel},
  issn         = {1948-7193},
  journal      = {ACS CHEMICAL NEUROSCIENCE},
  keywords     = {clozapine,5-HT7 receptor,olanzapine,GASP-1,lysosome,Helix-VIII,PROTEIN-COUPLED RECEPTORS,4TH CYTOPLASMIC LOOP,FUNCTIONAL SELECTIVITY,SORTING PROTEINS,INTRACELLULAR TRAFFICKING,ADENYLYL-CYCLASE,SPLICE VARIANTS,DRUG DISCOVERY,D2 RECEPTORS,EFFICACY},
  language     = {eng},
  number       = {7},
  pages        = {1206--1218},
  title        = {Downregulation of 5-HT7 serotonin receptors by the atypical antipsychotics clozapine and olanzapine: role of motifs in the C-terminal domain and interaction with GASP-1},
  url          = {http://dx.doi.org/10.1021/cn500339p},
  volume       = {6},
  year         = {2015},
}

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