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

Ivanova, Irina M., Nepogodiev, Sergey A., Saalbach, Gerhard, O'Neill, Ellis C., Urbaniak, Michael D., Ferguson, Michael A. J., Gurcha, Sudagar S., Besra, Gurdyal S. and Field, Robert A. ORCID: https://orcid.org/0000-0001-8574-0275 (2017) Fluorescent mannosides serve as acceptor substrates for glycosyltransferase and sugar-1-phosphate transferase activities in Euglena gracilis membranes. Carbohydrate Research, 438. pp. 26-38. ISSN 0008-6215

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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.

Item Type:	Article
Additional Information:	Funding Information: These studies were supported by the UK BBSRC Institute Strategic Programme on Understanding and Exploiting Metabolism (MET) [ BB/J004561/1 ] and the John Innes Foundation . We thank Martin Rejzek for help with HPLC analyses and Sakonwan Kuhaudomlarp for assistance with protein sequence analysis. Publisher Copyright: © 2016 The Authors
Uncontrolled Keywords:	enzyme assays,euglena gracilis,fluorescent glycans,glycosyltransferases,n-acetylglucosamine-1-phosphate transferase,analytical chemistry,biochemistry,organic chemistry ,/dk/atira/pure/subjectarea/asjc/1600/1602
Faculty \ School:	Faculty of Science > School of Chemistry, Pharmacy and Pharmacology
Related URLs:	http://www.scopus.com/inward/record.url?...
Depositing User:	LivePure Connector
Date Deposited:	04 Sep 2024 13:34
Last Modified:	02 Oct 2024 10:30
URI:	https://ueaeprints.uea.ac.uk/id/eprint/96524
DOI:	10.1016/j.carres.2016.11.017

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