对氧磷酶催化N-酰基高丝氨酸内酯水解的高效液相色谱分析
High-performance liquid chromatography analysis of N-acyl homoserine lactone hydrolysis by paraoxonases.
作者信息
Teiber John F, Draganov Dragomir I
机构信息
Division of Epidemiology, Department of Internal Medicine, The University of Texas Southwestern Medical Center, Dallas, TX, USA.
出版信息
Methods Mol Biol. 2011;692:291-8. doi: 10.1007/978-1-60761-971-0_21.
Mammalian paraoxonases (PONs) are a unique, highly conserved family of calcium-dependent esterases consisting of PON1, PON2, and PON3. The PONs can hydrolyze the lactone ring of a range of N-acyl-L: -homoserine lactone (AHL) quorum sensing signaling molecules, rendering them inactive. This chapter describes a method that utilizes high-performance liquid chromatography analysis with UV detection for determining the rate of AHL hydrolysis in cell lysates, tissue homogenates, serum, and with purified proteins. Also described are the techniques used to prepare cell culture lysates and tissue homogenates for analysis and the use of class-specific enzyme inhibitors to determine the contribution of PONs to AHL hydrolysis in the samples.
哺乳动物对氧磷酶（PONs）是一个独特的、高度保守的钙依赖性酯酶家族，由PON1、PON2和PON3组成。PONs可以水解一系列N-酰基-L-高丝氨酸内酯（AHL）群体感应信号分子的内酯环，使其失去活性。本章介绍了一种利用高效液相色谱分析和紫外检测来测定细胞裂解物、组织匀浆、血清以及纯化蛋白中AHL水解速率的方法。还介绍了用于制备细胞培养裂解物和组织匀浆以进行分析的技术，以及使用类特异性酶抑制剂来确定PONs对样品中AHL水解作用的方法。
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