TY - JOUR
T1 - Fluorescent mannosides serve as acceptor substrates for glycosyltransferase and sugar-1-phosphate transferase activities in Euglena gracilis membranes
AU - Ivanova, Irina M
AU - Nepogodiev, Sergey A
AU - Saalbach, Gerhard
AU - O'Neill, Ellis C
AU - Urbaniak, Michael D
AU - Ferguson, Michael A J
AU - Gurcha, Sudagar S
AU - Besra, Gurdyal S
AU - Field, Robert A
N1 - Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.
PY - 2017/1/13
Y1 - 2017/1/13
N2 - 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.
AB - 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.
KW - Euglena gracilis
KW - Glycosyltransferases
KW - Fluorescent glycans
KW - N-acetylglucosamine-1-phosphate transferase
KW - Enzyme assays
U2 - 10.1016/j.carres.2016.11.017
DO - 10.1016/j.carres.2016.11.017
M3 - Article
C2 - 27960097
SN - 0008-6215
VL - 438
SP - 26
EP - 38
JO - Carbohydrate Research
JF - Carbohydrate Research
ER -