title: Assay of Monoacylglycerol Lipase Activity.
abstract: Monoacylglycerol lipase (MGL) is a serine hydrolase involved in the biological deactivation of the endocannabinoid 2-arachidonoyl-sn-glycerol (2-AG). 2-AG is one of the main 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 helps control 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 the reaction product. This method may be used to determine the basal or altered MGL activity in various cells or animal tissues after pharmacological, genetic, or biological manipulations. In addition, this assay can be used for MGL inhibitor screening using purified recombinant enzyme or MGL-overexpressing cells.
authors.affiliation: Department of Anatomy and Neurobiology, University of California, Irvine, Irvine, CA, 92697, USA. kmjung@uci.edu.
authors.forename: Kwang-Mook
authors.initials: KM
authors.lastname: Jung
authors.affiliation: Department of Anatomy and Neurobiology, University of California, Irvine, Irvine, CA, 92697, USA.|Department of Pharmacology, University of California, Irvine, Irvine, CA, 92697, USA.|Department of Biological Chemistry, University of California, Irvine, Irvine, CA, 92697, USA.|Drug Discovery and Development, Istituto Italiano di Tecnologia, 16163, Genoa, Italy.
authors.forename: Daniele
authors.initials: D
authors.lastname: Piomelli
mesh_terms: D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D001095:Arachidonic Acids / Q000378:metabolism; D002384:Catalysis; D002460:Cell Line; D002853:Chromatography, Liquid; D063388:Endocannabinoids / Q000378:metabolism; D004789:Enzyme Activation; D057075:Enzyme Assays; D005989:Glycerides / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry; D008994:Monoacylglycerol Lipases / Q000378:metabolism*
chemical_list: D001095:Arachidonic Acids; D063388:Endocannabinoids; D005989:Glycerides; D016718:Arachidonic Acid; C094503:glyceryl 2-arachidonate; D008994:Monoacylglycerol Lipases
keywords: 2-Arachidonoyl-sn-glycerol (2-AG); Arachidonic acid; Enzyme assay; Liquid chromatography/mass spectrometry (LC/MS); Monoacylglycerol lipase (MGL)