Endocannabinoids Anandamide and 2-Arachidonoylglycerol Are Substrates for Human CYP2J2 Epoxygenase
| Autor: | Aditi Das, Amogh Kambalyal, Daryl D. Meling, Daniel R. McDougle |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Male
Epoxygenase Polyunsaturated Alkamides Swine Stereochemistry 2-Arachidonoylglycerol Arachidonic Acids Cytochrome P-450 CYP2J2 Metabolism Transport and Pharmacogenomics Protein Structure Secondary Glycerides Substrate Specificity CYP2J2 chemistry.chemical_compound Cytochrome P-450 Enzyme System Animals Humans CYP2C8 Pharmacology biology Cytochrome P450 reductase Cytochrome P450 Anandamide Protein Structure Tertiary Rats chemistry Biochemistry biology.protein Molecular Medicine Cattle Female lipids (amino acids peptides and proteins) Arachidonic acid Endocannabinoids Protein Binding |
| Zdroj: | Journal of Pharmacology and Experimental Therapeutics. 351:616-627 |
| ISSN: | 1521-0103 0022-3565 |
| DOI: | 10.1124/jpet.114.216598 |
| Popis: | The endocannabinoids, anandamide (AEA) and 2-arachidonoylglycerol (2-AG), are arachidonic acid (AA) derivatives that are known to regulate human cardiovascular functions. CYP2J2 is the primary cytochrome P450 in the human heart and is most well known for the metabolism of AA to the biologically active epoxyeicosatrienoic acids. In this study, we demonstrate that both 2-AG and AEA are substrates for metabolism by CYP2J2 epoxygenase in the model membrane bilayers of nanodiscs. Reactions of CYP2J2 with AEA formed four AEA-epoxyeicosatrienoic acids, whereas incubations with 2-AG yielded detectable levels of only two 2-AG epoxides. Notably, 2-AG was shown to undergo enzymatic oxidative cleavage to form AA through a NADPH-dependent reaction with CYP2J2 and cytochrome P450 reductase. The formation of the predominant AEA and 2-AG epoxides was confirmed using microsomes prepared from the left myocardium of porcine and bovine heart tissues. The nuances of the ligand-protein interactions were further characterized using spectral titrations, stopped-flow small-molecule ligand egress, and molecular modeling. The experimental and theoretical data were in agreement, which showed that substitution of the AA carboxylic acid with the 2-AG ester-glycerol changes the binding interaction of these lipids within the CYP2J2 active site, leading to different product distributions. In summary, we present data for the functional metabolomics of AEA and 2-AG by a membrane-bound cardiovascular epoxygenase. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|