Thumser A E, Voysey J, Wilton D C
Department of Biochemistry, University of Southampton, United Kingdom.
Biochem Pharmacol. 1997 Feb 7;53(3):433-5. doi: 10.1016/s0006-2952(96)00720-4.
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.
我们描述了一种简单的荧光位移测定法，用于测量两种重要的药理化合物花生四烯酰乙醇胺和油酰酰胺的水解。这些配体酰胺键处的水解会释放出一种脂肪酸作为产物之一。因此，释放出的脂肪酸会将大鼠肝脏脂肪酸结合蛋白上的荧光脂肪酸类似物取代，荧光的降低可用于测量这一过程。该过程具有时间和浓度依赖性，并呈现双曲线酶动力学。采用电喷雾电离质谱法对该测定法进行验证。
