Complex N-Glycans Influence the Spatial Arrangement of Voltage Gated Potassium Channels in Membranes of Neuronal-Derived Cells

PloS One
M Kristen HallRuth A Schwalbe

Abstract

The intrinsic electrical properties of a neuron depend on expression of voltage gated potassium (Kv) channel isoforms, as well as their distribution and density in the plasma membrane. Recently, we showed that N-glycosylation site occupancy of Kv3.1b modulated its placement in the cell body and neurites of a neuronal-derived cell line, B35 neuroblastoma cells. To extrapolate this mechanism to other N-glycosylated Kv channels, we evaluated the impact of N-glycosylation occupancy of Kv3.1a and Kv1.1 channels. Western blots revealed that wild type Kv3.1a and Kv1.1 α-subunits had complex and oligomannose N-glycans, respectively, and that abolishment of the N-glycosylation site(s) generated Kv proteins without N-glycans. Total internal reflection fluorescence microscopy images revealed that N-glycans of Kv3.1a contributed to its placement in the cell membrane while N-glycans had no effect on the distribution of Kv1.1. Based on particle analysis of EGFP-Kv proteins in the adhered membrane, glycosylated forms of Kv3.1a, Kv1.1, and Kv3.1b had differences in the number, size or density of Kv protein clusters in the cell membrane of neurites and cell body of B35 cells. Differences were also observed between the unglycosylated forms of th...Continue Reading

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Citations

Jun 16, 2016·International Journal of Molecular Sciences·M Kristen HallRuth A Schwalbe
Jun 9, 2018·The Journal of General Physiology·Angelica Lopez-Rodriguez, Miguel Holmgren
Jun 15, 2018·PloS One·M Kristen HallRuth A Schwalbe
Jun 10, 2021·Molecular Omics·Inga MatthiesMartin R Larsen

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Methods Mentioned

BETA
transfection
FACS
acetylation
glycosylation
protein folding

Software Mentioned

^ TIRF Control
Metamorph for
Cell
Olympus Basic
Image J

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