A high-throughput-compatible assay for determining the activity of fatty acid amide hydrolase.
Fatty acid amide hydrolase (EC 3.5.1.4.) is the enzyme responsible for the rapid degradation of lipid-derived chemical messengers such as anandamide, oleamide, and 2-arachidonoylglycerol. The pharmacological characterization of this enzyme in vivo has been hampered by the lack of selective and bioavailable inhibitors. We have developed a simple, radioactive, high-throughput-compatible assay for this enzyme based on the differential absorption of the substrate and its products to activated charcoal. The assay was validated using known inhibitors. It may be applied for the identification of new inhibitors from a compound library.
Wilson
Sandy J
SJ
Johnson & Johnson Pharmaceutical Research & Development LLC Neuroscience, 3210 Merryfield Row, San Diego, CA 92121, USA.
Lovenberg
Timothy W
TW
Barbier
Ann J
AJ
D000042:Absorption; D000581:Amidohydrolases / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000378:metabolism*; D001095:Arachidonic Acids / Q000737:chemistry / Q000378:metabolism; D045744:Cell Line, Tumor; D002606:Charcoal; D063388:Endocannabinoids; D004791:Enzyme Inhibitors / Q000302:isolation & purification / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D015394:Molecular Structure; D053284:Polyunsaturated Alkamides; D011851:Radioactivity; D012015:Reference Standards; D015203:Reproducibility of Results; D000099252:Fatty Acid Amide Hydrolases
D000581:Amidohydrolases; D001095:Arachidonic Acids; D002606:Charcoal; D063388:Endocannabinoids; D004791:Enzyme Inhibitors; D053284:Polyunsaturated Alkamides; D000099252:Fatty Acid Amide Hydrolases; C078814:anandamide