荧光法测定 FAAH 活性。

Fluorimetric Assay of FAAH Activity.

机构信息

Faculty of Veterinary Medicine, University of Teramo, Teramo, Italy.

Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, L'Aquila, Italy.

出版信息

Methods Mol Biol. 2023;2576:249-260. doi: 10.1007/978-1-0716-2728-0_21.

DOI:10.1007/978-1-0716-2728-0_21

PMID:36152193

Abstract

Fatty acid amide hydrolase (FAAH) is the enzyme responsible for the degradation of anandamide (N-arachidonoylethanolamine, AEA) to arachidonic acid (AA) and ethanolamine. The method described here measures FAAH activity through the fluorometric arachidonoyl-7-amino-4-methyl-coumarin amide (AAMCA) substrate, which allows a simple and sensitive assay suitable for high-throughput screening tests. FAAH catalyzes the hydrolysis of AAMCA producing AA and the highly fluorescent compound 7-amino-4-methylcoumarin (AMC).

摘要

脂肪酸酰胺水解酶（FAAH）是负责降解花生四烯酸乙醇胺（N-花生四烯酸乙醇胺，AEA）生成花生四烯酸（AA）和乙醇胺的酶。这里描述的方法通过荧光素酰基-7-氨基-4-甲基香豆素酰胺（AAMCA）底物来测量 FAAH 的活性，这使得该方法成为一种简单灵敏的测定方法，适用于高通量筛选试验。FAAH 催化 AAMCA 的水解生成 AA 和高荧光化合物 7-氨基-4-甲基香豆素（AMC）。