β-Arrestin1 and GPCR kinase2 play permissive roles in Src-mediated endocytosis of α4β2 nicotinic ACh receptors.
14-3-3
GRK2
Src
endocytosis
nicotine
α4β2 nAChR
β-arrestin1
Journal
British journal of pharmacology
ISSN: 1476-5381
Titre abrégé: Br J Pharmacol
Pays: England
ID NLM: 7502536
Informations de publication
Date de publication:
09 2021
09 2021
Historique:
revised:
02
04
2021
received:
07
10
2020
accepted:
06
04
2021
pubmed:
13
4
2021
medline:
22
9
2021
entrez:
12
4
2021
Statut:
ppublish
Résumé
The α4β2 nicotinic ACh receptor (nAChR), a subtype of the ligand-gated ion channel, is abundantly expressed in the brain and is implicated in several neurological disorders. The endocytosis of nAChRs plays important roles in the pathogenesis of neurological diseases, but the underlying molecular mechanisms remain poorly understood. Loss-of-function approaches and mutants of α4β2 nAChRs that display different endocytic properties were used to identify the cellular components and processes responsible for endocytosis. The signalling cascade that leads to endocytosis was deduced via protein interactions in predicted cellular components. The endocytosis of α4β2 nAChRs was determined and crosschecked using an ELISA and radioligand assay. Endocytosis of α4β2 nAChRs occurred through clathrin-mediated endocytosis in a dynamin-dependent manner. 14-3-3η-dependent Src-mediated phosphorylation of the nAChR α4 subunit at Y575 was required for nAChR endocytosis, and this occurred with the assistance of β-arrestin1 and GPCR kinase 2 (GRK2) without the need for kinase activity. Endocytosis triggered the mouse double minute 2 homologue-mediated ubiquitination and subsequent down-regulation of α4β2 nAChRs. α4β2 nAChR, an ionophore receptor, employs the metabotropic signalling pathway required for endocytosis, which leads to ubiquitination and down-regulation. Further, GRK2 and β-arrestin1, usually associated with GPCR signalling, are involved in the endocytosis of α4β2 nAChRs via different mechanisms. Considering the functional and pathological implications of nAChR endocytosis, results obtained in this study are crucial for the progression of basic research and clinical investigations.
Sections du résumé
BACKGROUND AND PURPOSE
The α4β2 nicotinic ACh receptor (nAChR), a subtype of the ligand-gated ion channel, is abundantly expressed in the brain and is implicated in several neurological disorders. The endocytosis of nAChRs plays important roles in the pathogenesis of neurological diseases, but the underlying molecular mechanisms remain poorly understood.
EXPERIMENTAL APPROACH
Loss-of-function approaches and mutants of α4β2 nAChRs that display different endocytic properties were used to identify the cellular components and processes responsible for endocytosis. The signalling cascade that leads to endocytosis was deduced via protein interactions in predicted cellular components. The endocytosis of α4β2 nAChRs was determined and crosschecked using an ELISA and radioligand assay.
KEY RESULTS
Endocytosis of α4β2 nAChRs occurred through clathrin-mediated endocytosis in a dynamin-dependent manner. 14-3-3η-dependent Src-mediated phosphorylation of the nAChR α4 subunit at Y575 was required for nAChR endocytosis, and this occurred with the assistance of β-arrestin1 and GPCR kinase 2 (GRK2) without the need for kinase activity. Endocytosis triggered the mouse double minute 2 homologue-mediated ubiquitination and subsequent down-regulation of α4β2 nAChRs.
CONCLUSIONS AND IMPLICATIONS
α4β2 nAChR, an ionophore receptor, employs the metabotropic signalling pathway required for endocytosis, which leads to ubiquitination and down-regulation. Further, GRK2 and β-arrestin1, usually associated with GPCR signalling, are involved in the endocytosis of α4β2 nAChRs via different mechanisms. Considering the functional and pathological implications of nAChR endocytosis, results obtained in this study are crucial for the progression of basic research and clinical investigations.
Substances chimiques
Receptors, Nicotinic
0
Acetylcholine
N9YNS0M02X
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
3498-3516Informations de copyright
© 2021 The British Pharmacological Society.
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