Lack of complex type N-glycans lessens aberrant neuronal properties

ECU Author/Contributor (non-ECU co-authors, if there are any, appear on document)

M. Kristen Hall (Creator)

Ruth A. Schwalbe (Creator)

Douglas A. Weidner (Creator)

Austin A. Whitman (Creator)

Institution

East Carolina University (ECU )

Web Site: http://www.ecu.edu/lib/

Abstract: Modifications in surface glycans attached to proteins via N-acetylglucosamine-ß1-Nasparagine linkage have been linked to tumor development and progression. These modifications include complex N-glycans with high levels of branching , fucose and sialic acid residues. Previously , we silenced Mgat2 in neuroblastoma (NB) cells , which halted the conversion of hybrid type N-glycans to complex type , to generate a novel cell line , NB_1 (-Mgat2). By comparing the aberrant cell properties of the NB_1(-Mgat2) cell line to the parental cell line (NB_1) , we investigated the impact of eliminating complex type N-glycans on NB cell behavior. Further , the N-glycosylation pathway in the NB_1(-Mgat2) cell line was rescued by transiently transfecting cells with Mgat2 , thus creating the NB_1 (-/+Mgat2) cell line. Changes in the N-glycosylation pathway were verified by enhanced binding of E-PHA and L-PHA to proteins in the rescued cell line relative to those of the NB_1(-Mgat2) cell line. Also , western blotting of total membranes from the rescued cell line ectopically expressing a voltage-gated K+ channel (Kv3.1b) revealed that N-glycans of Kv3.1b were processed to complex type. By employment of various cell lines , we demonstrated that reduction of the complex type N-glycans diminished anchorage-independent cell growth , and enhanced cell-cell interactions. Two independent cell invasion assays showed that cell invasiveness was markedly lessened by lowering the levels of complex type N-glycans while cell mobility was only slightly modified. Neurites of NB cells were shortened by the absence of complex type N-glycans. Cell proliferation was reduced in NB cells with lowered levels of complex type N-glycans which resulted from hindered progression through G1+Go phases of the cell cycle. Overall , our results illustrate that reducing the ratio of complex to hybrid types of N-glycans diminishes aberrant NB cell behavior and thereby has a suppressive effect in cell proliferation , and cell dissociation and invasion phases of NB.

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Hall , M. K. , Weidner , D. A. , Whitman , A. A. , & Schwalbe , R. A. (2018). Lack of complex type N-glycans lessens aberrant neuronal properties. PloS One , 13(6) , e0199202. doi:10.1371/journal.pone.0199202

Language: English

Date: 2018

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