N-glycosylation promotes the cell surface expression of Kv1.3 potassium channels

Jing Zhu, Jenny Yan, William B. Thornhill

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

The voltage-gated potassium channel Kv1.3 plays an essential role in modulating membrane excitability in many cell types. Kv1.3 is a heavily glycosylated membrane protein. Two successive N-glycosylation consensus sites, N228NS and N229ST, are present on the S1-S2 linker of rat Kv1.3. Our data suggest that Kv1.3 contains only one N-glycan and it is predominantly attached to N229 in the S1-S2 extracellular linker. Preventing N-glycosylation of Kv1.3 significantly decreased its surface protein level and surface conductance density level, which were ∼ 49% and ∼ 46% respectively of the level of wild type. Supplementation of N-acetylglucosamine (GlcNAc), l-fucose or N-acetylneuraminic acid to the culture medium promoted Kv1.3 surface protein expression, whereas supplementation of d-glucose, d-mannose or d-galactose did not. Among the three effective monosaccharides/derivatives, adding GlcNAc appeared to reduce sialic acid content and increase the degree of branching in the N-glycan of Kv1.3, suggesting that the N-glycan structure and composition had changed. Furthermore, the cell surface half-life of the Kv1.3 surface protein was increased upon GlcNAc supplementation, indicating that it had decreased internalization. The GlcNAc effect appears to apply mainly to membrane proteins containing complex type N-glycans. Thus, N-glycosylation promotes Kv1.3 cell surface expression; supplementation of GlcNAc increased Kv1.3 surface protein level and decreased its internalization, presumably by a combined effect of decreased branch size and increased branching of the N-glycan. N-glycosylation and supplementation with particular monosaccharides affected the surface expression of the Kv1.3 potassium channel. Preventing N-glycosylation decreased its surface protein and conductance levels by two-fold, whereas GlcNAc, l-fucose, or sialic acid in the culture medium increased its surface protein level by 1.5-3.0-fold. Thus the N-glycan and its branch size or degree of branching affected the surface levels of Kv1.3

Original languageEnglish (US)
Pages (from-to)2632-2644
Number of pages13
JournalFEBS Journal
Volume279
Issue number15
DOIs
StatePublished - Aug 2012
Externally publishedYes

Keywords

  • N-glycosylation
  • cell surface expression
  • monosaccharide supplementation
  • potassium channel
  • trafficking

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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