Project description:NanoString nCounter analysis using the human Host Response Panel was used to determine the expression of 618 genes involved in host response to infection from telomerase immortalized gingival keratinocytes (TIGKs) incubated with the oral pathogen Selenomonas sputigena ATCC 35185 compared to untreated control cells.
Project description:The Gram-negative, flagellated, anaerobic, crescent-shaped bacterium Selenomonas sputigena is a potential human periodontal pathogen. Information on its virulence factors and underlying pathogenicity mechanisms is scarce. Here we show for the first time that S. sputigena produces a diversely and heavily O-glycosylated flagellin C9LY14 as a major cellular protein, which carries various hitherto undescribed rhamnose- and N acetylglucosamine-linked O-glycans in the range from mono- to hexasaccharides. A comprehensive glycomic and glycoproteomic assessment revealed extensive glycan macro- and microheterogeneity on 20 unique glycopeptide species. From the multiple sites of glycosylation, five were unambiguously identified on the 437-amino acid C9LY14 protein (Thr149, Ser182, Thr199, Thr259, and Ser334). The O-glycans additionally showed modifications by methylation and acetylation. This is the first report on O-linked flagellin glycosylation in S. sputigena, revealing that C9LY14 is one of the most heavily glycosylated flagellins described to date.