Wheeler, Simon, Breen, Colum, Li, Yong, Hewitt, Sarah H, Robertson, Erin, Yates, Edwin A ORCID: 0000-0001-9365-5433, Barsukov, Igor L, Fernig, David G ORCID: 0000-0003-4875-4293 and Butler, Stephen J
(2021)
Anion binding to a cationic europium(iii) probe enables the first real-time assay of heparan sulfotransferase activity.
ORGANIC & BIOMOLECULAR CHEMISTRY, 20 (3).
pp. 596-605.
Abstract
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.
Item Type: | Article |
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Uncontrolled Keywords: | Anions, Cations, Europium, Sulfotransferases, Phosphoadenosine Phosphosulfate, Molecular Structure, Time Factors, Coordination Complexes |
Divisions: | Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology |
Depositing User: | Symplectic Admin |
Date Deposited: | 18 May 2022 09:09 |
Last Modified: | 18 Jan 2023 21:01 |
DOI: | 10.1039/d1ob02071d |
Open Access URL: | https://doi.org/10.1039/D1OB02071D |
Related URLs: | |
URI: | https://livrepository.liverpool.ac.uk/id/eprint/3155024 |