Assay of Monoacylglycerol Lipase Activity.
Assay of Monoacylglycerol Lipase Activity.
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单酰基甘油脂肪酶活性的测定。
DOI：
10.1007/978-1-0716-2728-0_24
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Piomelli,Daniele
中科院分区：
文献类型：
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作者：
Jung,Kwang-Mook;Piomelli,Daniele
单酰基甘油脂肪酶(MGL/MAGL/MGLL)是一种丝氨酸水解酶，参与了2-AG生物失活的过程。2-AG是大脑和身体其他部位最丰富的大麻素受体内源性脂质激动剂。在中枢神经系统中，MGL定位于兴奋性和抑制性突触的突触前神经末梢，控制2-AG对突触传递和可塑性的调节作用。在这一章中，我们描述了一种体外评估MGL活性的方法，该方法基于LC/MS对其反应产物进行定量。该方法可用于测定药物、遗传或生物干预后细胞或组织中的基础或改变的MGL活性。此外，该方法还可用于利用纯化的重组酶或MGL过表达细胞筛选MGL抑制剂。
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.