An optimised MALDI-TOF assay for phosphatidylcholine-specific phospholipase C.

Reynisson

doi:10.1039/D0AY02208J

An optimised MALDI-TOF assay for phosphatidylcholine-specific phospholipase C.

Reynisson

Authors

Johannes Reynisson j.reynisson@keele.ac.uk

Abstract

The Bacillus cereus phosphatidylcholine-specific phospholipase C (PC-PLCBc) is an enzyme that catalyses the hydrolysis of phosphatidylcholines into phosphocholine and 1,2-diacylglycerols. PC-PLCBc has found applications in both the food industry and in medicinal chemistry. Herein, we report our work in the development and optimisation of a matrix assisted laser desorption ionisation time-of-flight (MALDI-TOF) mass spectrometry-based assay to monitor PC-PLCBc activity. The use of one-phase and two-phase reaction systems to assess the inhibition of PC-PLCBc with different structural classes of inhibitors was compared. We also highlighted the advantage of our assay over the commonly used commercially available Amplex Red assay. This method will also be applicable to work on the activity and inhibition of other phospholipases.

Acceptance Date	Dec 29, 2020
Publication Date	Jan 12, 2021
Journal	Analytical Methods
Print ISSN	1759-9660
Publisher	Royal Society of Chemistry
DOI	https://doi.org/10.1039/D0AY02208J
Publisher URL	https://pubs.rsc.org/en/content/articlelanding/2021/AY/D0AY02208J#!divAbstract