A novel broad specificity fucosidase capable of core alpha 1-6 fucose release from N-glycans labeled with urea-linked fluorescent dyes

作者：Vainauskas Saulius; Kirk Charlotte H; Petralia Laudine; Guthrie Ellen P; McLeod Elizabeth; Bielik Alicia; Luebbers Alex; Foster Jeremy M; Hokke Cornelis H; Rudd Pauline M; Shi Xiaofeng; Taron Christopher H^*

来源：Scientific Reports, 2018, 8(1): 9504.

DOI：10.1038/s41598-018-27797-0

Exoglycosidases are often used for detailed characterization of glycan structures. Bovine kidney alpha-fucosidase is commonly used to determine the presence of core alpha 1-6 fucose on N-glycans, an important modification of glycoproteins. Recently, several studies have reported that removal of core alpha 1-6-linked fucose from N-glycans labeled with the reactive N-hydroxysuccinimide carbamate fluorescent labels 6-aminoquinolyl-N-hydroxysuccinimidylcarbamate (AQC) and RapiFluor-MS is severely impeded. We report here the cloning, expression and biochemical characterization of an alpha-fucosidase from Omnitrophica bacterium (termed fucosidase O). We show that fucosidase O can efficiently remove alpha 1-6- and alpha 1-3-linked core fucose from N-glycans. Additionally, we demonstrate that fucosidase O is able to efficiently hydrolyze core alpha 1-6-linked fucose from N-glycans labeled with any of the existing NHS-carbamate activated fluorescent dyes.

• 出版日期2018-6-22
• 单位New England Biolabs