Profiling of core fucosylated N-glycans using a novel bacterial lectin that specifically recognizes α1,6 fucosylated chitobiose.
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 α1-6 fucosylated N-glycans, but not to core α1-3 fucosylated N-glycans, or other α1-2, α1-3 and α1-4 fucosylated oligosaccharides. The minimal high affinity binding epitope of SL2-1 was α1-6 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.
New England Biolabs, 240 County Road, Ipswich, MA 01938, USA.
Saulius
S
Vainauskas
New England Biolabs, 240 County Road, Ipswich, MA 01938, USA.|Charles River Laboratories, 8 Henshaw St., Woburn, MA 01801, USA.
Rebecca M
RM
Duke
New England Biolabs, 240 County Road, Ipswich, MA 01938, USA.
James
J
McFarland
New England Biolabs, 240 County Road, Ipswich, MA 01938, USA.
Colleen
C
McClung
New England Biolabs, 240 County Road, Ipswich, MA 01938, USA.
Cristian
C
Ruse
New England Biolabs, 240 County Road, Ipswich, MA 01938, USA.
Christopher H
CH
Taron
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