Assay of Endocannabinoid Oxidation by Cyclooxygenase-2.
Abstract
The endocannabinoids, 2-arachidonoylglycerol (2-AG) and arachidonylethanolamide (AEA), are endogenous ligands for the cannabinoid receptors (CB1 and CB2) and are implicated in a wide array of physiological processes. These neutral arachidonic acid (AA) derivatives have been identified as efficient substrates for the second isoform of the cyclooxygenase enzyme (COX-2). A diverse family of prostaglandin glycerol esters (PG-Gs) and prostaglandin ethanolamides (PG-EAs) is generated by the action of COX-2 (and downstream prostaglandin synthases) on 2-AG and AEA. As the biological importance of the endocannabinoid system becomes more apparent, there is a tremendous need for robust, sensitive, and efficient analytical methodology for the endocannabinoids and their metabolites. In this chapter, we describe methodology suitable for carrying out oxygenation of endocannabinoids by COX-2, and analysis of products of endocannabinoid oxygenation by COX-2 and of endocannabinoids themselves from in vitro and cell assays.
A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Vanderbilt Institute of Chemical Biology, Vanderbilt Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, 37232, USA. Department of Biochemistry, Vanderbilt Institute of Chemical Biology, Vanderbilt Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, 37232, USA.A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Vanderbilt Institute of Chemical Biology, Vanderbilt Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, 37232, USA. Department of Biochemistry, Vanderbilt Institute of Chemical Biology, Vanderbilt Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, 37232, USA. Department of Chemistry, Vanderbilt Institute of Chemical Biology, Vanderbilt Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, 37232, USA.A.B. Hancock Jr. Memorial Laboratory for Cancer Research, Vanderbilt Institute of Chemical Biology, Vanderbilt Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, 37232, USA. larry.marnett@vanderbilt.edu. Department of Biochemistry, Vanderbilt Institute of Chemical Biology, Vanderbilt Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, 37232, USA. larry.marnett@vanderbilt.edu. Department of Chemistry, Vanderbilt Institute of Chemical Biology, Vanderbilt Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, 37232, USA. larry.marnett@vanderbilt.edu. Department of Pharmacology, Vanderbilt Institute of Chemical Biology, Vanderbilt Center in Molecular Toxicology, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, 37232, USA. larry.marnett@vanderbilt.edu. MeSH
AnimalsArachidonic AcidArachidonic AcidsBiological AssayCell LineChromatography, LiquidCyclooxygenase 2EndocannabinoidsGlyceridesIn Vitro TechniquesMacrophagesMiceOxidation-ReductionPolyunsaturated AlkamidesSubstrate SpecificityTandem Mass Spectrometry
Pub Type(s)
Journal Article
Research Support, N.I.H., Extramural
