Profiling of core fucosylated N-glycans using a novel bacterial lectin that specifically recognizes ?1,6 fucosylated chitobiose.

ABSTRACT: A novel fucose-binding lectin (SL2-1) from the bacterium Streptomyces rapamycinicus was identified by analysis of metagenomic DNA sequences. SL2-1 belongs to a new group of bacterial fucose-specific lectins that have no similarity to known bacterial fucose-binding proteins, but are related to certain eukaryotic fucose-binding lectins. The 17?kDa protein was expressed recombinantly in E. coli and purified by affinity chromatography. Glycan microarray analysis with fluorescently labeled recombinant SL2-1 demonstrated its ability to bind to core ?1-6 fucosylated N-glycans, but not to core ?1-3 fucosylated N-glycans, or other ?1-2, ?1-3 and ?1-4 fucosylated oligosaccharides. The minimal high affinity binding epitope of SL2-1 was ?1-6 fucosylated di-n-acetylchitobiose. The recombinant lectin was efficient in detection of N-glycan core fucosylation using lectin blotting and lectin ELISA assays. Finally, a workflow using SL2-1 for selective and quantitative profiling of core fucosylated N-glycans using UPLC-HILIC-FLR analysis was established. The approach was validated for selective capture and analysis of core fucosylated N-glycans present in complex glycan mixtures derived from mammalian serum IgG.

SUBMITTER: Vainauskas S 

PROVIDER: S-EPMC5039751 | biostudies-literature | 2016 Sep

REPOSITORIES: biostudies-literature