Cargando…

Distinct Cell Surface Expression Patterns of N-Glycosylation Site Mutants of AMPA-Type Glutamate Receptor under the Homo-Oligomeric Expression Conditions

The AMPA-type glutamate receptor (AMPAR) is a homotetrameric or heterotetrameric ion channel composed of various combinations of four subunits (GluA1–4), and its abundance in the synapse determines the strength of synaptic activity. The formation of oligomers in the endoplasmatic reticulum (ER) is c...

Descripción completa

Detalles Bibliográficos
Autores principales: Morise, Jyoji, Yamamoto, Saki, Midorikawa, Ryosuke, Takamiya, Kogo, Nonaka, Motohiro, Takematsu, Hiromu, Oka, Shogo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7404163/
https://www.ncbi.nlm.nih.gov/pubmed/32707739
http://dx.doi.org/10.3390/ijms21145101
_version_ 1783567091436093440
author Morise, Jyoji
Yamamoto, Saki
Midorikawa, Ryosuke
Takamiya, Kogo
Nonaka, Motohiro
Takematsu, Hiromu
Oka, Shogo
author_facet Morise, Jyoji
Yamamoto, Saki
Midorikawa, Ryosuke
Takamiya, Kogo
Nonaka, Motohiro
Takematsu, Hiromu
Oka, Shogo
author_sort Morise, Jyoji
collection PubMed
description The AMPA-type glutamate receptor (AMPAR) is a homotetrameric or heterotetrameric ion channel composed of various combinations of four subunits (GluA1–4), and its abundance in the synapse determines the strength of synaptic activity. The formation of oligomers in the endoplasmatic reticulum (ER) is crucial for AMPAR subunits’ ER-exit and translocation to the cell membrane. Although N-glycosylation on different AMPAR subunits has been shown to regulate the ER-exit of hetero-oligomers, its role in the ER-exit of homo-oligomers remains unclear. In this study, we investigated the role of N-glycans at GluA1N63/N363 and GluA2N370 in ER-exit under the homo-oligomeric expression conditions, whose mutants are known to show low cell surface expressions. In contrast to the N-glycosylation site mutant GluA1N63Q, the cell surface expression levels of GluA1N363Q and GluA2N370Q increased in a time-dependent manner. Unlike wild-type (WT) GluA1, GluA2WT rescued surface GluA2N370Q expression. Additionally, the expression of GluA1N63Q reduced the cell surface expression level of GluA1WT. In conclusion, our findings suggest that these N-glycans have distinct roles in the ER-exit of GluA1 and GluA2 homo-oligomers; N-glycan at GluA1N63 is a prerequisite for GluA1 ER-exit, whereas N-glycans at GluA1N363 and GluA2N370 control the ER-exit rate.
format Online
Article
Text
id pubmed-7404163
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-74041632020-08-11 Distinct Cell Surface Expression Patterns of N-Glycosylation Site Mutants of AMPA-Type Glutamate Receptor under the Homo-Oligomeric Expression Conditions Morise, Jyoji Yamamoto, Saki Midorikawa, Ryosuke Takamiya, Kogo Nonaka, Motohiro Takematsu, Hiromu Oka, Shogo Int J Mol Sci Article The AMPA-type glutamate receptor (AMPAR) is a homotetrameric or heterotetrameric ion channel composed of various combinations of four subunits (GluA1–4), and its abundance in the synapse determines the strength of synaptic activity. The formation of oligomers in the endoplasmatic reticulum (ER) is crucial for AMPAR subunits’ ER-exit and translocation to the cell membrane. Although N-glycosylation on different AMPAR subunits has been shown to regulate the ER-exit of hetero-oligomers, its role in the ER-exit of homo-oligomers remains unclear. In this study, we investigated the role of N-glycans at GluA1N63/N363 and GluA2N370 in ER-exit under the homo-oligomeric expression conditions, whose mutants are known to show low cell surface expressions. In contrast to the N-glycosylation site mutant GluA1N63Q, the cell surface expression levels of GluA1N363Q and GluA2N370Q increased in a time-dependent manner. Unlike wild-type (WT) GluA1, GluA2WT rescued surface GluA2N370Q expression. Additionally, the expression of GluA1N63Q reduced the cell surface expression level of GluA1WT. In conclusion, our findings suggest that these N-glycans have distinct roles in the ER-exit of GluA1 and GluA2 homo-oligomers; N-glycan at GluA1N63 is a prerequisite for GluA1 ER-exit, whereas N-glycans at GluA1N363 and GluA2N370 control the ER-exit rate. MDPI 2020-07-19 /pmc/articles/PMC7404163/ /pubmed/32707739 http://dx.doi.org/10.3390/ijms21145101 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Morise, Jyoji
Yamamoto, Saki
Midorikawa, Ryosuke
Takamiya, Kogo
Nonaka, Motohiro
Takematsu, Hiromu
Oka, Shogo
Distinct Cell Surface Expression Patterns of N-Glycosylation Site Mutants of AMPA-Type Glutamate Receptor under the Homo-Oligomeric Expression Conditions
title Distinct Cell Surface Expression Patterns of N-Glycosylation Site Mutants of AMPA-Type Glutamate Receptor under the Homo-Oligomeric Expression Conditions
title_full Distinct Cell Surface Expression Patterns of N-Glycosylation Site Mutants of AMPA-Type Glutamate Receptor under the Homo-Oligomeric Expression Conditions
title_fullStr Distinct Cell Surface Expression Patterns of N-Glycosylation Site Mutants of AMPA-Type Glutamate Receptor under the Homo-Oligomeric Expression Conditions
title_full_unstemmed Distinct Cell Surface Expression Patterns of N-Glycosylation Site Mutants of AMPA-Type Glutamate Receptor under the Homo-Oligomeric Expression Conditions
title_short Distinct Cell Surface Expression Patterns of N-Glycosylation Site Mutants of AMPA-Type Glutamate Receptor under the Homo-Oligomeric Expression Conditions
title_sort distinct cell surface expression patterns of n-glycosylation site mutants of ampa-type glutamate receptor under the homo-oligomeric expression conditions
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7404163/
https://www.ncbi.nlm.nih.gov/pubmed/32707739
http://dx.doi.org/10.3390/ijms21145101
work_keys_str_mv AT morisejyoji distinctcellsurfaceexpressionpatternsofnglycosylationsitemutantsofampatypeglutamatereceptorunderthehomooligomericexpressionconditions
AT yamamotosaki distinctcellsurfaceexpressionpatternsofnglycosylationsitemutantsofampatypeglutamatereceptorunderthehomooligomericexpressionconditions
AT midorikawaryosuke distinctcellsurfaceexpressionpatternsofnglycosylationsitemutantsofampatypeglutamatereceptorunderthehomooligomericexpressionconditions
AT takamiyakogo distinctcellsurfaceexpressionpatternsofnglycosylationsitemutantsofampatypeglutamatereceptorunderthehomooligomericexpressionconditions
AT nonakamotohiro distinctcellsurfaceexpressionpatternsofnglycosylationsitemutantsofampatypeglutamatereceptorunderthehomooligomericexpressionconditions
AT takematsuhiromu distinctcellsurfaceexpressionpatternsofnglycosylationsitemutantsofampatypeglutamatereceptorunderthehomooligomericexpressionconditions
AT okashogo distinctcellsurfaceexpressionpatternsofnglycosylationsitemutantsofampatypeglutamatereceptorunderthehomooligomericexpressionconditions