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Anion binding to a cationic europium(iii) probe enables the first real-time assay of heparan sulfotransferase activity

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journal contribution
posted on 13.04.2022, 12:50 by Simon WheelerSimon Wheeler, Colum Breen, Yong Li, Sarah Hewitt, Erin Robertson, Edwin A Yates, Igor L Barsukov, David G Fernig, Stephen ButlerStephen Butler

Sulfotransferases constitute a ubiquitous class of enzymes which are poorly understood due to the lack of a convenient tool for screening their activity. These enzymes use the anion PAPS (adenosine-3′-phosphate-5′-phosphosulfate) as a donor for a broad range of acceptor substrates, including carbohydrates, producing sulfated compounds and PAP (adenosine-3′,5′-diphosphate) as a side product. We present a europium(III)-based probe that binds reversibly to both PAPS and PAP, producing a larger luminescence enhancement with the latter anion. We exploit this greater emission enhancement with PAP to demonstrate the first direct real-time assay of a heparan sulfate sulfotransferase using a multi-well plate format. The selective response of our probe towards PAP over structurally similar nucleoside phosphate anions, and over other anions, is investigated and discussed. This work opens the possibility of investigating more fully the roles played by this enzyme class in health and disease, including operationally simple inhibitor screening.

Funding

Biotechnology and Biotechnological Sciences Research Council Tools and Resources Development Grant [BB/T012099/1]

European Commission FET-OPEN grant [ArrestAD no.737390]

Biotechnology and Biotechnological Sciences Research Council Industrial Biotechnology [BB/V003372/1]

History

School

  • Science

Department

  • Chemistry

Published in

Organic & Biomolecular Chemistry

Volume

20

Issue

3

Pages

596 - 605

Publisher

Royal Society of Chemistry (RSC)

Version

VoR (Version of Record)

Rights holder

Journal is © The Royal Society of Chemistry

Publisher statement

This is an Open Access Article. It is published by the Royal Society of Chemistry under the Creative Commons Attribution 3.0 Unported Licence (CC BY 3.0). Full details of this licence are available at: https://creativecommons.org/licenses/by/3.0/

Acceptance date

18/11/2021

Publication date

2021-12-08

Copyright date

2021

ISSN

1477-0520

eISSN

1477-0539

Language

en

Depositor

Deposit date: 11 April 2022