An optimised MALDI-TOF assay for phosphatidylcholine-specific phospholipase C,Analytical Methods

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An optimised MALDI-TOF assay for phosphatidylcholine-specific phospholipase C
Analytical Methods ( IF 2.7 ) Pub Date : 2020-12-29 , DOI: 10.1039/d0ay02208j
Nabangshu Sharma _{1,

2,

3,

4} , Ries J. Langley _{2,

3,

4,

5,

6} , Chatchakorn Eurtivong _{7,

8,

9,

10,

11} , Euphemia Leung _{2,

3,

4,

5,

6} , Ryan Joseph Dixon _{1,

2,

3,

4} , Emily K. Paulin _{1,

2,

3,

4} , Shaun W. P. Rees _{1,

2,

3,

4} , Lisa I. Pilkington _{1,

2,

3,

4} , David Barker _{1,

2,

3,

4,

12} , Jóhannes Reynisson _{1,

2,

3,

4,

13} , Ivanhoe K. H. Leung _{1,

2,

3,

4,

5}

Affiliation

The Bacillus cereus phosphatidylcholine-specific phospholipase C (PC-PLC_Bc) is an enzyme that catalyses the hydrolysis of phosphatidylcholines into phosphocholine and 1,2-diacylglycerols. PC-PLC_Bc 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-PLC_Bc activity. The use of one-phase and two-phase reaction systems to assess the inhibition of PC-PLC_Bc 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.

中文翻译：

磷脂酰胆碱特异性磷脂酶C的优化MALDI-TOF测定

的蜡状芽孢杆菌的磷脂酰胆碱特异性磷脂酶C（PC-PLC _BC）是一种酶，其催化磷脂酰胆碱水解成磷酸胆碱和1,2-二酰基甘油。PC-PLC _Bc已在食品工业和药物化学中得到应用。在此，我们报告了我们在基于基质辅助激光解吸电离飞行时间（MALDI-TOF）质谱的检测方法的开发和优化中的工作，以监测PC-PLC _Bc的活性。使用一相和两相反应系统评估对PC-PLC _Bc的抑制比较了具有不同结构类别的抑制剂。我们还强调了我们的检测方法相对于常用的市售Amplex Red检测方法的优势。该方法也可用于研究其他磷脂酶的活性和抑制作用。

更新日期：2021-01-12

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