A novel broad specificity fucosidase capable of core α1-6 fucose release from N-glycans labeled with urea-linked fluorescent dyes

  • Sci Rep. 2018 Jun 22;8(1):9504. doi: 10.1038/s41598-018-27797-0.
Saulius Vainauskas  1 ,  Charlotte H Kirk  1 ,  Laudine Petralia  1  2 ,  Ellen P Guthrie  1 ,  Elizabeth McLeod  1 ,  Alicia Bielik  1 ,  Alex Luebbers  1 ,  Jeremy M Foster  1 ,  Cornelis H Hokke  2 ,  Pauline M Rudd  3 ,  Xiaofeng Shi  1 ,  Christopher H Taron  4
Affiliations
  • 1. New England Biolabs, 240 County Road, Ipswich, MA, 01938, USA.
  • 2. Department of Parasitology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA, Leiden, The Netherlands.
  • 3. NIBRT GlycoScience Group, National Institute for Bioprocessing, Research and Training, Foster's Avenue, Mount Merrion, Blackrock, Co, Dublin, Ireland.
  • 4. New England Biolabs, 240 County Road, Ipswich, MA, 01938, USA. [email protected].
PMID: 29934601 DOI: 10.1038/s41598-018-27797-0
Abstract

Exoglycosidases are often used for detailed characterization of glycan structures. Bovine kidney α-fucosidase is commonly used to determine the presence of core α1-6 fucose on N-glycans, an important modification of glycoproteins. Recently, several studies have reported that removal of core α1-6-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 α-fucosidase from Omnitrophica bacterium (termed fucosidase O). We show that fucosidase O can efficiently remove α1-6- and α1-3-linked core fucose from N-glycans. Additionally, we demonstrate that fucosidase O is able to efficiently hydrolyze core α1-6-linked fucose from N-glycans labeled with any of the existing NHS-carbamate activated fluorescent dyes.

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