Fluorescent mannosides serve as acceptor substrates for glycosyltransferase and sugar-1-phosphate transferase activities in Euglena gracilis membranes

Irina M. Ivanova, Sergey A. Nepogodiev, Gerhard Saalbach, Ellis C. O'Neill, Michael D. Urbaniak, Michael A. J. Ferguson, Sudagar S. Gurcha, Gurdyal S. Besra, Robert A. Field (Lead / Corresponding author)

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8 Citations (Scopus)
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Abstract

Synthetic hexynyl α-D-mannopyranoside and its α-1,6-linked disaccharide counterpart were fluorescently labelled through CuAAC click chemistry with 3-azido-7-hydroxycoumarin. The resulting triazolyl-coumarin adducts, which were amenable to analysis by TLC, HPLC and mass spectrometry, proved to be acceptor substrates for α-1,6-ManT activities in mycobacterial membranes, as well as α- and β-GalT activities in trypanosomal membranes, benchmarking the potential of the fluorescent acceptor approach against earlier radiochemical assays. Following on to explore the glycobiology of the benign protozoan alga Euglena gracilis, α-1,3- and α-1,2-ManT activities were detected in membrane preparations, along with GlcT, Glc-P-T and GlcNAc-P-T activities. These studies serve to demonstrate the potential of readily accessible fluorescent glycans as substrates for exploring carbohydrate active enzymes.

Original languageEnglish
Pages (from-to)26-38
Number of pages13
JournalCarbohydrate Research
Volume438
Early online date30 Nov 2016
DOIs
Publication statusPublished - 13 Jan 2017

Keywords

  • Euglena gracilis
  • Glycosyltransferases
  • Fluorescent glycans
  • N-acetylglucosamine-1-phosphate
  • transferase
  • Enzyme assays

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