用于磷脂酰胆碱特异性磷脂酶 C 的优化 MALDI-TOF 分析。
An optimised MALDI-TOF assay for phosphatidylcholine-specific phospholipase C.
机构信息
School of Chemical Sciences, The University of Auckland, Private Bag 92019, Victoria Street West, Auckland 1142, New Zealand.
Maurice Wilkins Centre for Molecular Biodiscovery, The University of Auckland, Private Bag 92019, Victoria Street West, Auckland 1142, New Zealand and Department of Molecular Medicine and Pathology, School of Medical Sciences, The University of Auckland, Private Bag 92019, Victoria Street West, Auckland 1142, New Zealand.
出版信息
Anal Methods. 2021 Feb 4;13(4):491-496. doi: 10.1039/d0ay02208j.
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.
蜡状芽孢杆菌磷脂酰胆碱特异性磷脂酶 C（PC-PLCBc）是一种酶，能够催化磷脂酰胆碱水解为磷酸胆碱和 1,2-二酰基甘油。PC-PLCBc 在食品工业和药物化学领域都有应用。本文报道了我们在开发和优化基质辅助激光解吸电离飞行时间（MALDI-TOF）质谱检测法以监测 PC-PLCBc 活性方面的工作。比较了单相和两相反应系统在评估不同结构类别抑制剂对 PC-PLCBc 抑制作用方面的应用。我们还强调了该方法相对于常用的商业可用的 Amplex Red 测定法的优势。这种方法也适用于研究其他磷脂酶的活性和抑制作用。
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