Journal article
An optimised MALDI-TOF assay for phosphatidylcholine-specific phospholipase C
N Sharma, RJ Langley, C Eurtivong, E Leung, RJ Dixon, EK Paulin, SWP Rees, LI Pilkington, D Barker, J Reynisson, IKH Leung
Analytical Methods | ROYAL SOC CHEMISTRY | Published : 2021
DOI: 10.1039/d0ay02208j
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 commer..
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Awarded by Health Research Council of New Zealand
Funding Acknowledgements
We thank Dr Praju Vikas Anekal of the University of Auckland for providing training and technical support for MALDI-TOF mass spectrometry. We thank the generous financial support from the University of Auckland, the Cancer Society of New Zealand (Ref: 17.01), a Breast Cancer Research Partnership project grant from the Health Research Council of New Zealand, Breast Cancer Cure, and Breast Cancer Foundation NZ (Ref: 17/671), a Ronald Kay Science Fellowship from Breast Cancer Foundation NZ (L. I. P.) and a Rutherford Foundation Postdoctoral Fellowship from the Royal Society Te Aparangi-Rutherford Foundation (L. I. P.).