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

Department of Biological Sciences and Center for Cancer, Genetic Diseases and Gene Regulation, Fordham University, Bronx, NY, USA.
FEBS Journal (Impact Factor: 4). 05/2012; 279(15):2632-44. DOI: 10.1111/j.1742-4658.2012.08642.x
Source: PubMed

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.

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    • "e for K v 1 . 4 where desialylation has no impact on the channel gating , in contrast to K v 1 . 1 and K v 1 . 5 ( Watanabe et al . 2003 ) . Functional studies on K v 1 . 2 and K v 1 . 3 also revealed that preventing glycosylation drastically alters surface trafficking ( increased ER retention ) and gating of the channels ( Watanabe et al . 2007 ; Zhu et al . 2012 ) ."
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