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Assay of Endocannabinoid Oxidation by Cyclooxygenase-2.
Methods Mol Biol. 2016; 1412:205-15.MM

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.

Authors+Show Affiliations

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.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

27245906

Citation

Kudalkar, Shalley N., et al. "Assay of Endocannabinoid Oxidation By Cyclooxygenase-2." Methods in Molecular Biology (Clifton, N.J.), vol. 1412, 2016, pp. 205-15.
Kudalkar SN, Kingsley PJ, Marnett LJ. Assay of Endocannabinoid Oxidation by Cyclooxygenase-2. Methods Mol Biol. 2016;1412:205-15.
Kudalkar, S. N., Kingsley, P. J., & Marnett, L. J. (2016). Assay of Endocannabinoid Oxidation by Cyclooxygenase-2. Methods in Molecular Biology (Clifton, N.J.), 1412, 205-15. https://doi.org/10.1007/978-1-4939-3539-0_21
Kudalkar SN, Kingsley PJ, Marnett LJ. Assay of Endocannabinoid Oxidation By Cyclooxygenase-2. Methods Mol Biol. 2016;1412:205-15. PubMed PMID: 27245906.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Assay of Endocannabinoid Oxidation by Cyclooxygenase-2. AU - Kudalkar,Shalley N, AU - Kingsley,Philip J, AU - Marnett,Lawrence J, PY - 2016/6/2/entrez PY - 2016/6/2/pubmed PY - 2017/12/8/medline KW - Cell assay KW - Cyclooxygenase-2 KW - Endocannabinoids KW - In vitro assay KW - LC-MS/MS KW - PG-EAs KW - PG-Gs SP - 205 EP - 15 JF - Methods in molecular biology (Clifton, N.J.) JO - Methods Mol Biol VL - 1412 N2 - 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. SN - 1940-6029 UR - https://www.unboundmedicine.com/medline/citation/27245906/Assay_of_Endocannabinoid_Oxidation_by_Cyclooxygenase_2_ L2 - https://dx.doi.org/10.1007/978-1-4939-3539-0_21 DB - PRIME DP - Unbound Medicine ER -