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Profiling substrate promiscuity of wild-type sugar kinases for multifluorinated monosaccharides

Profiling substrate promiscuity of wild-type sugar kinases for multifluorinated monosaccharides
Profiling substrate promiscuity of wild-type sugar kinases for multifluorinated monosaccharides
Fluorinated sugar-1-phosphates are of emerging importance as intermediates in the chemical and biocatalytic synthesis of modified oligosaccharides, as well as probes for chemical biology. Here we present a systematic study of the activity of a wide range of anomeric sugar kinases (galacto- and N-acetylhexosamine kinases) against a panel of fluorinated monosaccharides, leading to the first examples of polyfluorinated substrates accepted by this class of enzymes. We have discovered four new N-acetylhexosamine kinases with a different substrate scope, thus expanding the number of homologs available in this subclass of kinases. Lastly, we have solved the crystal structure of a galactokinase in complex with 2-deoxy-2fluoro galactose, giving insight into changes in the active site that may account for the specificity of the enzyme towards certain substrate analogues.
biocatalysis, kinases, sugar phosphates, fluorinated carbohydrates, oligosaccharides, glycobiology, enzyme discovery
2451-9448
1199-1206.e5
Keenan, Tessa
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Parmeggiani, Fabio
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Malassis, Julien GJ
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Fontenelle, Clement Q
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Vendeville, Jean B
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Offen, Wendy
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Both, Peter
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Huang, Kun
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Marchesi, Andrea
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Heyam, Alex
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Young, Carl
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Charnock, Simon J.
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Davies, Gideon J.
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Linclau, Bruno
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Flitsch, Sabine L.
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Fascione, Martin A.
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Keenan, Tessa
2480c679-d88c-421b-b697-188a5fa60bcd
Parmeggiani, Fabio
61347984-e2cd-404f-b9ab-3521ef5c5a14
Malassis, Julien GJ
5cd4224d-e4db-4835-863c-56b709511ccd
Fontenelle, Clement Q
c746c5ea-96bc-46c2-849d-47215ab58c03
Vendeville, Jean B
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Offen, Wendy
ddc1a5c1-04bb-460b-a4cd-0c3acc4a9b9a
Both, Peter
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Huang, Kun
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Marchesi, Andrea
3cc57216-aedc-48ba-a7f3-0464393fd3cc
Heyam, Alex
fb273457-ed16-4a60-843f-e9e9ca8b5fb6
Young, Carl
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Charnock, Simon J.
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Davies, Gideon J.
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Linclau, Bruno
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Flitsch, Sabine L.
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Fascione, Martin A.
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Keenan, Tessa, Parmeggiani, Fabio, Malassis, Julien GJ, Fontenelle, Clement Q, Vendeville, Jean B, Offen, Wendy, Both, Peter, Huang, Kun, Marchesi, Andrea, Heyam, Alex, Young, Carl, Charnock, Simon J., Davies, Gideon J., Linclau, Bruno, Flitsch, Sabine L. and Fascione, Martin A. (2020) Profiling substrate promiscuity of wild-type sugar kinases for multifluorinated monosaccharides. Cell Chemical Biology, 27 (9), 1199-1206.e5. (doi:10.1016/j.chembiol.2020.06.005).

Record type: Article

Abstract

Fluorinated sugar-1-phosphates are of emerging importance as intermediates in the chemical and biocatalytic synthesis of modified oligosaccharides, as well as probes for chemical biology. Here we present a systematic study of the activity of a wide range of anomeric sugar kinases (galacto- and N-acetylhexosamine kinases) against a panel of fluorinated monosaccharides, leading to the first examples of polyfluorinated substrates accepted by this class of enzymes. We have discovered four new N-acetylhexosamine kinases with a different substrate scope, thus expanding the number of homologs available in this subclass of kinases. Lastly, we have solved the crystal structure of a galactokinase in complex with 2-deoxy-2fluoro galactose, giving insight into changes in the active site that may account for the specificity of the enzyme towards certain substrate analogues.

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MAF_Kinases paper - _Cell_Chem_Biol_postprint - Accepted Manuscript
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Accepted/In Press date: 12 June 2020
e-pub ahead of print date: 2 July 2020
Published date: 17 September 2020
Keywords: biocatalysis, kinases, sugar phosphates, fluorinated carbohydrates, oligosaccharides, glycobiology, enzyme discovery

Identifiers

Local EPrints ID: 450595
URI: http://eprints.soton.ac.uk/id/eprint/450595
ISSN: 2451-9448
PURE UUID: 6b94184e-3493-494b-9492-654c51742508
ORCID for Clement Q Fontenelle: ORCID iD orcid.org/0000-0002-1630-3407
ORCID for Bruno Linclau: ORCID iD orcid.org/0000-0001-8762-0170

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Date deposited: 04 Aug 2021 16:35
Last modified: 17 Mar 2024 05:40

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Contributors

Author: Tessa Keenan
Author: Fabio Parmeggiani
Author: Julien GJ Malassis
Author: Clement Q Fontenelle ORCID iD
Author: Jean B Vendeville
Author: Wendy Offen
Author: Peter Both
Author: Kun Huang
Author: Andrea Marchesi
Author: Alex Heyam
Author: Carl Young
Author: Simon J. Charnock
Author: Gideon J. Davies
Author: Bruno Linclau ORCID iD
Author: Sabine L. Flitsch
Author: Martin A. Fascione

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