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High throughput mass spectrometry-based characterisation of Arabidopsis thaliana group H glycosyltransferases

Akere, A; Liu, Q; Wu, S; Hou, B; Yang, M; (2018) High throughput mass spectrometry-based characterisation of Arabidopsis thaliana group H glycosyltransferases. RSC Advances , 8 (53) pp. 30080-30086. 10.1039/c8ra03947j. Green open access

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Abstract

In this report, we cloned and characterised four members of group H glycosyltransferases (GTs) by studying their substrate specificities and kinetics. The formation of products and possible glycosylation position was confirmed using MS/MS. The results revealed that 76E1 and 76E5 have broader donor specificity, including UDP-glucose (UDPGlc), UDP-galactose (UDPGal) and UDP-N-acetylglucosamine (UDPGlcNAc) with various flavonoids as acceptor substrates. Pseudo-single substrate kinetics data showed a relatively low K_M, indicating a high affinity for substrate UDPGlc and also supported that 76E5 is more of a galactosyl and N-acetylglucosamine transferase. Sequence alignment and site-directed mutagenesis studies indeed suggested that serine is a crucial residue in the UDPGlcNAc and UDPGal activity.

Type: Article
Title: High throughput mass spectrometry-based characterisation of Arabidopsis thaliana group H glycosyltransferases
Open access status: An open access version is available from UCL Discovery
DOI: 10.1039/c8ra03947j
Publisher version: http://dx.doi.org/10.1039/c8ra03947j
Language: English
Additional information: © Royal Society of Chemistry 2018. This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence (https://creativecommons.org/licenses/by-nc/3.0/).
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > UCL School of Pharmacy
URI: https://discovery.ucl.ac.uk/id/eprint/10056201
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