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β1- and β3- voltage-gated sodium channel subunits modulate cell surface expression and glycosylation of Na(v)1.7 in HEK293 cells

Voltage-gated sodium channels (Na(v)s) are glycoproteins composed of a pore-forming α-subunit and associated β-subunits that regulate Na(v) α-subunit plasma membrane density and biophysical properties. Glycosylation of the Na(v) α-subunit also directly affects Na(v)s gating. β-subunits and glycosyla...

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Autores principales: Laedermann, Cédric J., Syam, Ninda, Pertin, Marie, Decosterd, Isabelle, Abriel, Hugues
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3757325/
https://www.ncbi.nlm.nih.gov/pubmed/24009557
http://dx.doi.org/10.3389/fncel.2013.00137
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author Laedermann, Cédric J.
Syam, Ninda
Pertin, Marie
Decosterd, Isabelle
Abriel, Hugues
author_facet Laedermann, Cédric J.
Syam, Ninda
Pertin, Marie
Decosterd, Isabelle
Abriel, Hugues
author_sort Laedermann, Cédric J.
collection PubMed
description Voltage-gated sodium channels (Na(v)s) are glycoproteins composed of a pore-forming α-subunit and associated β-subunits that regulate Na(v) α-subunit plasma membrane density and biophysical properties. Glycosylation of the Na(v) α-subunit also directly affects Na(v)s gating. β-subunits and glycosylation thus comodulate Na(v) α-subunit gating. We hypothesized that β-subunits could directly influence α-subunit glycosylation. Whole-cell patch clamp of HEK293 cells revealed that both β1- and β3-subunits coexpression shifted V(½) of steady-state activation and inactivation and increased Na(v)1.7-mediated I(Na) density. Biotinylation of cell surface proteins, combined with the use of deglycosydases, confirmed that Na(v)1.7 α-subunits exist in multiple glycosylated states. The α-subunit intracellular fraction was found in a core-glycosylated state, migrating at ~250 kDa. At the plasma membrane, in addition to the core-glycosylated form, a fully glycosylated form of Na(v)1.7 (~280 kDa) was observed. This higher band shifted to an intermediate band (~260 kDa) when β1-subunits were coexpressed, suggesting that the β1-subunit promotes an alternative glycosylated form of Na(v)1.7. Furthermore, the β1-subunit increased the expression of this alternative glycosylated form and the β3-subunit increased the expression of the core-glycosylated form of Na(v)1.7. This study describes a novel role for β1- and β3-subunits in the modulation of Na(v)1.7 α-subunit glycosylation and cell surface expression.
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spelling pubmed-37573252013-09-05 β1- and β3- voltage-gated sodium channel subunits modulate cell surface expression and glycosylation of Na(v)1.7 in HEK293 cells Laedermann, Cédric J. Syam, Ninda Pertin, Marie Decosterd, Isabelle Abriel, Hugues Front Cell Neurosci Neuroscience Voltage-gated sodium channels (Na(v)s) are glycoproteins composed of a pore-forming α-subunit and associated β-subunits that regulate Na(v) α-subunit plasma membrane density and biophysical properties. Glycosylation of the Na(v) α-subunit also directly affects Na(v)s gating. β-subunits and glycosylation thus comodulate Na(v) α-subunit gating. We hypothesized that β-subunits could directly influence α-subunit glycosylation. Whole-cell patch clamp of HEK293 cells revealed that both β1- and β3-subunits coexpression shifted V(½) of steady-state activation and inactivation and increased Na(v)1.7-mediated I(Na) density. Biotinylation of cell surface proteins, combined with the use of deglycosydases, confirmed that Na(v)1.7 α-subunits exist in multiple glycosylated states. The α-subunit intracellular fraction was found in a core-glycosylated state, migrating at ~250 kDa. At the plasma membrane, in addition to the core-glycosylated form, a fully glycosylated form of Na(v)1.7 (~280 kDa) was observed. This higher band shifted to an intermediate band (~260 kDa) when β1-subunits were coexpressed, suggesting that the β1-subunit promotes an alternative glycosylated form of Na(v)1.7. Furthermore, the β1-subunit increased the expression of this alternative glycosylated form and the β3-subunit increased the expression of the core-glycosylated form of Na(v)1.7. This study describes a novel role for β1- and β3-subunits in the modulation of Na(v)1.7 α-subunit glycosylation and cell surface expression. Frontiers Media S.A. 2013-08-30 /pmc/articles/PMC3757325/ /pubmed/24009557 http://dx.doi.org/10.3389/fncel.2013.00137 Text en Copyright © 2013 Laedermann, Syam, Pertin, Decosterd and Abriel. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Neuroscience
Laedermann, Cédric J.
Syam, Ninda
Pertin, Marie
Decosterd, Isabelle
Abriel, Hugues
β1- and β3- voltage-gated sodium channel subunits modulate cell surface expression and glycosylation of Na(v)1.7 in HEK293 cells
title β1- and β3- voltage-gated sodium channel subunits modulate cell surface expression and glycosylation of Na(v)1.7 in HEK293 cells
title_full β1- and β3- voltage-gated sodium channel subunits modulate cell surface expression and glycosylation of Na(v)1.7 in HEK293 cells
title_fullStr β1- and β3- voltage-gated sodium channel subunits modulate cell surface expression and glycosylation of Na(v)1.7 in HEK293 cells
title_full_unstemmed β1- and β3- voltage-gated sodium channel subunits modulate cell surface expression and glycosylation of Na(v)1.7 in HEK293 cells
title_short β1- and β3- voltage-gated sodium channel subunits modulate cell surface expression and glycosylation of Na(v)1.7 in HEK293 cells
title_sort β1- and β3- voltage-gated sodium channel subunits modulate cell surface expression and glycosylation of na(v)1.7 in hek293 cells
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3757325/
https://www.ncbi.nlm.nih.gov/pubmed/24009557
http://dx.doi.org/10.3389/fncel.2013.00137
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