Cell surface glycans remodeled in vitro using commercial glycosidases

MAGNELLI, PAULA; ALEX LUEBBERS; CAGNONI, ALEJANDRO J; MOROSI, LUCIANO; CUTINE, ANABELA; WANDS, AMBERLYN; KOHLER, JENNIFER; MARIÑO, KARINA

San Francisco, USA

Congreso; Society for Glycobiology Meeting; 2015

Society of Glycobiology

Glycan-protein interactions are involved in different biological processes including development, immunity, inflammation and cancer. As a result, manipulation of cell surface glycans is an essential tool for understanding the mechanisms by which glycans govern cell fate. Traditional approaches using gene KOs, gene silencing, or metabolic blockers are not always adequate (metabolic inhibitors have pleiotropic effects, glycosylation mutants are not available for every cell type).Specific glycosidases that effectively remove selected glycan residues at the cell surface could be a viable alternative to remodel the cell surface glycome when other methods are not suitable. Although this approach has been reported in the literature, there has never been a systematic study (comparing different cell lines, enzymes, and reaction conditions) where the extent and efficacy of glycan removal was evaluated along with a measurement of cell integrity and viability.We present studies showing the activity of a cocktail of exoglycosidase/s under conditions compatible with cell integrity. Based on these results, efficient in vitro cell surface glycan remodeling was possible, maintaining culture viability. After enzymatic treatment, modifications in the glycan structure were analyzed by flow cytometry using specific lectins.