Predominant Expression of Hybrid N-Glycans Has Distinct Cellular Roles Relative to Complex and Oligomannose N-Glycans
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2016-06
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Authors
Hall, M. Kristen
Weidner, Douglas A.
Zhu, Young
Dayal, Sahil
Whitman, Austin A.
Schwalbe, Ruth
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Abstract
Glycosylation modulates growth, maintenance, and stress signaling processes.
Consequently, altered N-glycosylation is associated with reduced fitness and disease. Therefore,
expanding our understanding of N-glycans in altering biological processes is of utmost interest.
Herein, clustered regularly interspaced short palindromic repeats/caspase9 (CRISPR/Cas9)
technology was employed to engineer a glycosylation mutant Chinese Hamster Ovary (CHO) cell line,
K16, which expresses predominantly hybrid type N-glycans. This newly engineered cell line enabled
us to compare N-glycan effects on cellular properties of hybrid type N-glycans, to the well-established
Pro´5 and Lec1 cell lines, which express complex and oligomannose types of N-glycans, respectively.
Lectin binding studies revealed the predominant N-glycan expressed in K16 is hybrid type. Cell
dissociation and migration assays demonstrated the greatest strength of cell–cell adhesion and fastest
migratory rates for oligomannose N-glycans, and these properties decreased as oligomannose type
were converted to hybrid type, and further decreased upon conversion to complex type. Next,
we examined the roles of three general types of N-glycans on ectopic expression of E-cadherin,
a cell–cell adhesion protein. Microscopy revealed more functional E-cadherin at the cell–cell border
when N-glycans were oligomannose and these levels decreased as the oligomannose N-glycans were
processed to hybrid and then to complex. Thus, we provide evidence that all three general types of
N-glycans impact plasma membrane architecture and cellular propertie
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DOI
10.3390/ijms17060925