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A fluorescence displacement assay for the measurement of arachidonoyl ethanolamide (anandamide) and oleoyl amide (octadecenoamide) hydrolysis

Authors
  • Thumser, Alfred E.A.
  • Voysey, Joanne
  • Wilton, David C.
Type
Published Article
Journal
Biochemical Pharmacology
Publisher
Elsevier
Publication Date
Jan 01, 1997
Accepted Date
Sep 10, 1996
Volume
53
Issue
3
Pages
433–435
Identifiers
DOI: 10.1016/S0006-2952(96)00720-4
Source
Elsevier
Keywords
License
Unknown

Abstract

We describe a simple fluorescence displacement assay to measure hydrolysis of arachidonoyl ethanolamide and oleoyl amide, two important pharmacological compounds. Hydrolysis at the amide linkage of these ligands releases a fatty acid as one of the products. The displacement of a fluorescent fatty acid analogue from rat liver fatty acid-binding protein by the released fatty acid can thus be measured as a decrease in fluorescence. This process is time- and concentration-dependent and shows hyperbolic enzyme kinetics. Electrospray ionisation mass spectrometry was used to validate the assay.

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