Douglas A Weidner
Tanzania
Research Article
Membrane Distribution and Activity of a Neuronal Voltage-Gated K+ Channel is Modified by Replacement of Complex Type N-Glycans with Hybrid Type
Author(s): M Kristen Hall, Douglas A Weidner, Sahil Dayal, Elena Pak, Alexander K Murashov and Ruth A SchwalbeM Kristen Hall, Douglas A Weidner, Sahil Dayal, Elena Pak, Alexander K Murashov and Ruth A Schwalbe
Abnormal modifications in N-glycosylation processing are commonly associated with neurological disorders, although the impact of specific N-glycans on neuronal excitability is unknown. By replacement of complex types of N-glycans with hybrid types in neuroblastoma cells, we provide the first study that addresses how distinct N-glycan types impact neuronal excitability. Using CRISPR/Cas9 technology, NB_1, a clonal cell line derived from rat neuroblastoma cells (NB), was modified to create an N-glycosylation mutant cell line, NB_1 (-Mgat2), which expresses predominantly hybrid type N-glycans. Western and lectin blotting, flow cytometry, TIRF and DIC microscopy, and patch clamp studies were conducted. Lectin binding revealed the predominant type of N-glycans expressed in NB_1 (-Mgat2) is hybrid while those of NB and NB_1 are complex. Kv3.1b-expressing cells with complex N-glycans localiz.. View More»
