磷脂磷酸酶(PAP)催化三酰甘油合成中的倒数第二步,并调节膜磷脂的合成。人们对这种酶非常感兴趣,因为它控制其底物的细胞水平,磷脂酸(PA),和产品,DAG;这些脂质中间体的代谢缺陷是肥胖等脂质疾病的基础,脂肪营养不良,和炎症。旨在了解其作用机制的研究需要测量PAP活性,它是如何被监管的,以及用于筛选其活化剂和/或抑制剂。通过使用测量产品的放射性和非放射性测定法来确定酶活性,DAG,或者Pi然而,在这些方法中,灵敏度和易用性是可变的。这篇综述总结了合成放射性PA的方法,分析放射性和非放射性产品,DAG和Pi,并讨论了每种PAP测定法的优缺点。
Phosphatidate phosphatase (PAP) catalyzes the penultimate step in the synthesis of triacylglycerol and regulates the synthesis of membrane phospholipids. There is much interest in this enzyme because it controls the cellular levels of its substrate, phosphatidate (PA), and product, DAG; defects in the metabolism of these lipid intermediates are the basis for lipid-based diseases such as obesity, lipodystrophy, and inflammation. The measurement of PAP activity is required for studies aimed at understanding its mechanisms of action, how it is regulated, and for screening its activators and/or inhibitors. Enzyme activity is determined through the use of radioactive and nonradioactive assays that measure the product, DAG, or Pi However, sensitivity and ease of use are variable across these methods. This
review summarizes approaches to synthesize radioactive PA, to analyze radioactive and nonradioactive products, DAG and Pi, and discusses the advantages and disadvantages of each PAP assay.