测定单酰甘油脂肪酶活性。

Assay of Monoacylglycerol Lipase Activity.

机构信息

Department of Anatomy and Neurobiology, University of California, Irvine, Irvine, CA, USA.

Department of Pharmacology, University of California, Irvine, Irvine, CA, USA.

出版信息

Methods Mol Biol. 2023;2576:285-297. doi: 10.1007/978-1-0716-2728-0_24.

DOI:10.1007/978-1-0716-2728-0_24

PMID:36152196

Abstract

Monoacylglycerol lipase (MGL/MAGL/MGLL) is a serine hydrolase involved in the biological deactivation of the endocannabinoid 2-arachidonoyl-sn-glycerol (2-AG). 2-AG is the most abundant endogenous lipid agonists for cannabinoid receptors in the brain and elsewhere in the body. In the central nervous system (CNS), MGL is localized to presynaptic nerve terminals of both excitatory and inhibitory synapses, where it controls the regulatory actions of 2-AG on synaptic transmission and plasticity. In this chapter, we describe an in vitro method to assess MGL activity by liquid chromatography/mass spectrometry (LC/MS)-based quantitation of its reaction product. The method may be used to determine basal or altered MGL activity in cells or tissues after pharmacological, genetic, or biological interventions. In addition, the assay can be used for MGL inhibitor screening using purified recombinant enzyme or MGL-overexpressing cells.

摘要

单酰甘油脂肪酶（MGL/MAGL/MGLL）是一种丝氨酸水解酶，参与生物失活内源性大麻素 2-花生四烯酰基-sn-甘油（2-AG）。2-AG 是大脑和身体其他部位中大麻素受体的最丰富的内源性脂质激动剂。在中枢神经系统（CNS）中，MGL 定位于兴奋性和抑制性突触的突触前神经末梢，在那里它控制 2-AG 对突触传递和可塑性的调节作用。在本章中，我们描述了一种基于液相色谱/质谱（LC/MS）的体外方法，通过定量分析其反应产物来评估 MGL 活性。该方法可用于在药理学、遗传学或生物学干预后测定细胞或组织中的基础或改变的 MGL 活性。此外，该测定法还可用于使用纯化的重组酶或过表达 MGL 的细胞进行 MGL 抑制剂筛选。