{Reference Type}: Journal Article
{Title}: A fluorogenic probe for core-fucosylated glycan-preferred ENGase.
{Author}: Ishii N;Muto H;Nagata M;Sano K;Sato I;Iino K;Matsuzaki Y;Katoh T;Yamamoto K;Matsuo I;
{Journal}: Carbohydr Res
{Volume}: 523
{Issue}: 0
{Year}: Jan 2023
{Factor}: 2.975
{DOI}: 10.1016/j.carres.2022.108724
{Abstract}: A fluorescence-quenching-based assay system to determine the hydrolytic activity of endo-β-N-acetylglucosaminidases (ENGases), which act on the innermost N-acetylglucosamine (GlcNAc) residue of the chitobiose segment of core-fucosylated N-glycans, was constructed using a dual-labeled fluorescent probe with a hexasaccharide structure. The fluorogenic probe was evaluated using a variety of ENGases, including Endo-M W251N mutant, Endo-F3, and Endo-S, which recognize core fucosylated N-glycans. The occurrence of a hydrolysis reaction was detected by observing an increased fluorescence intensity, ultimately allowing the ENGase activities to be easily and quantitatively evaluated, with the exception of Endo-S. The obtained results clearly indicated the substrate specificities of the examined ENGases.