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

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9 Citations (Scopus)
161 Downloads (Pure)

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
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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