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

Research output: Contribution to journalArticle

Authors

  • Irina M Ivanova
  • Sergey A Nepogodiev
  • Gerhard Saalbach
  • Ellis C O'Neill
  • Michael D Urbaniak
  • Michael A J Ferguson
  • Robert A Field

Colleges, School and Institutes

External organisations

  • Department of Biological Chemistry, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, UK.
  • Biomedical and Life Sciences, Lancaster University, Furness Building, Lancaster LA1 4YG, UK; College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
  • University of Dundee
  • Department of Biological Chemistry, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, UK. Electronic address: rob.field@jic.ac.uk.

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.

Details

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

Keywords

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