Moser, Sandra;
Hans, Gloria;
Shukla, Anuj;
Saiardi, Adolfo;
Bru, Samuel;
Taskin, Asli Aras;
Meisinger, Chris;
(2025)
A screening approach unveils an unknown Mn2+-dependent endopolyphosphatase activity in yeast.
Chemical Science
, 16
(30)
pp. 13760-13768.
10.1039/d5sc02166a.
Preview |
Text
A screening approach unveils an unknown Mn<sup>2+<sup>-dependent endopolyphosphatase activity in yeast.pdf - Published Version Download (1MB) | Preview |
Abstract
Inorganic polyphosphate (polyP) is a ubiquitous biopolymer composed of multiple orthophosphates connected by energy-rich phosphoanhydride bonds. In organisms, polyP is digested by two types of enzymes: exopolyphosphatases, which shorten the chain from the ends by cleaving off monophosphate units, and endopolyphosphatases, which cut the chain internally. While several continuous methods are available to monitor exopolyphosphatase activity, endopolyphosphatase activity assays are less common and typically involve multiple tedious steps. Here, we introduce FRET-polyP8, a novel probe for real-time detection of endopolyphosphatase activity. The FRET assay enabled rapid, highly sensitive, single-step detection of specific endopolyphosphatase activity both from isolated proteins and cell extracts. The simple read-out additionally enabled enzyme inhibitor screening. Furthermore, a novel Mn2+-dependent endopolyphosphatase activity in baker's yeast was detected in a quadruple mutant, highlighting the ability to screen for metal-dependence of new endopolyphosphatase activity. This approach thus represents a significant addition to existing methodologies, facilitating the discovery and classification of new endopolyphosphatases and their inhibitors to advance our understanding of polyP metabolism and regulation.
| Type: | Article |
|---|---|
| Title: | A screening approach unveils an unknown Mn2+-dependent endopolyphosphatase activity in yeast |
| Location: | England |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1039/d5sc02166a |
| Publisher version: | https://doi.org/10.1039/d5sc02166a |
| Language: | English |
| Additional information: | This article is licensed under a Creative Commons Attribution 3.0 Unported Licence, https://creativecommons.org/licenses/by/3.0/. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given. |
| 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 > Lab for Molecular Cell Bio MRC-UCL |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10215063 |
Archive Staff Only
 |
View Item |
