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Cited by 1Glycosylation and Glycoproteins ResearchIon channel regulation and functionProtein Structure and Dynamics

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Structural implications of glycosylation on the voltage-gated sodium channel β3-subunit

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Beaudoin, Christopher A., Hayes, Jerrard, Liu, Hengrui, Namadurai, Sivakumar, Deery, Michael J., August, Ethan A., Krasner, Samuel, Hamaia, Samir W., Salvage, Samantha C., Huang, Christopher L.-H., Jackson, Antony P.·2026·Journal of Structural Biology
DOI 10.1016/j.jsb.2026.108321

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Abstract

Abstract Voltage-gated sodium (Na V ) channel α-subunits are modulated by associated β-subunits that affect their localization, trafficking and gating behaviour. The β-subunits are members of the immunoglobulin (Ig) domain family of cell-adhesion molecules and the interactions between their extracellular Ig-domains may modify channel clustering. The full-length β3-subunit can form cis trimers on the plasma membrane. The atomic resolution structure of a deglycosylated trimeric β3-subunit Ig-domain has been solved by X-ray crystallography. However, it is not clear whether this particular trimeric Ig-domain structure is plausible for cell-expressed, glycosylated β3-subunits. Here we use glycan profiling to confirm an extensive and heterogeneous pattern of β3-subunit glycosylation, with the majority of glycans being bi- and tri-antennary structures with one or two terminal sialic acids. Two tryptic peptides of the β3 Ig-domain are predicted to contain potential N-linked glycosylation sites. When the isolated, glycosylated full-length β3-subunit was trypsin-digested and analysed by LC-MS/MS, only one of these peptides - containing an N-linked glycosylation site at residue N95 and located close to the trimer interface - was identified in its unmodified form, suggesting that residue N95 is under-glycosylated. All-atom molecular dynamics simulations of the glycosylated, membrane-bound full-length β3 trimer confirmed that glycans can be accommodated with the Ig-domain trimer and indeed, may contribute to protein-membrane and inter-protomer interactions within the full-length, membrane-embedded trimer. Further biochemical studies are warranted to explore the interactions between oligomeric β-subunits with corresponding α-subunit sodium channels.

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Metadata

Authors
Beaudoin, C. A., Hayes, J., Liu, H., Namadurai, S., Deery, M. J., August, E. A., Krasner, S., Hamaia, S. W., Salvage, S. C., Huang, C. L.-H., & Jackson, A. P.
Affiliations
  • University of Cambridge (GB)
  • Trinity College Dublin (IE)
  • John Radcliffe Hospital (GB)
  • St George's, University of London (GB)
Corresponding author
  • Beaudoin, Christopher A.
  • Jackson, Antony P.
ORCID
Venue
Journal of Structural Biology
Publisher
Elsevier BV
Published
2026-06
Volume
218
Issue
2
Pages
108321
ISSN
1047-8477 (ISSN-L)
DOI
10.1016/j.jsb.2026.108321
OpenAlex
W4408007185
Cited by
1
References
1
Citations by year
2026: 1
FWCI
2.09
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Structural Biology (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • SSRN Electronic Journal (acceptedVersion)
  • PubMed (publishedVersion)
  • St George's Online Research Archive (St George's University of London) (acceptedVersion)
  • Apollo (University of Cambridge) (submittedVersion, cc-by)
Funders
  • University of Cambridge
  • British Heart Foundation (PG/24/12121, PG/19/59/34582, RE/24/130024)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion channel regulation and function
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
Keywords
voltage-gated sodium channels, beta3 subunit, glycosylation, immunoglobulin domains, N-glycosylation sites, sialic acid, protein-membrane interactions, channel clustering, glycan profiling, molecular dynamics simulation
MeSH terms
Immunoglobulin Domains, Animals, Glycosylation, Humans, Models, Molecular, Polysaccharides, Crystallography, X-Ray, Protein Subunits, Voltage-Gated Sodium Channels

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