Journal article

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

Nabangshu Sharma, Ries J Langley, Chatchakorn Eurtivong, Euphemia Leung, Ryan Joseph Dixon, Emily K Paulin, Shaun WP Rees, Lisa Pilkington, David Barker, Johannes Reynisson, Ivanhoe KH Leung

ANALYTICAL METHODS | ROYAL SOC CHEMISTRY | Published : 2021

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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University of Melbourne Researchers

Grants

Awarded by Cancer Society of New Zealand


Awarded by Breast Cancer Foundation NZ


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.).