title: Assay of NAAA Activity.
abstract: N-acylethanolamine-hydrolyzing acid amidase (NAAA) is a lysosomal hydrolase degrading various N-acylethanolamines at acidic pH. Since NAAA prefers anti-inflammatory and analgesic palmitoylethanolamide to other N-acylethanolamines as a substrate, its specific inhibitors are expected as a new class of anti-inflammatory and analgesic agents. Here, we introduce an NAAA assay system, using [(14)C]palmitoylethanolamide and thin-layer chromatography. The preparation of NAAA enzyme from native and recombinant sources as well as the chemical synthesis of N-[1'-(14)C]palmitoyl-ethanolamine is also described.
authors.affiliation: Department of Biochemistry, Kagawa University School of Medicine, 1750-1 Ikenobe, Miki, Kagawa, 761-0793, Japan.
authors.forename: Kazuhito
authors.initials: K
authors.lastname: Tsuboi
authors.affiliation: Department of Biochemistry, Kagawa University School of Medicine, 1750-1 Ikenobe, Miki, Kagawa, 761-0793, Japan. nueda@med.kagawa-u.ac.jp.
authors.forename: Natsuo
authors.initials: N
authors.lastname: Ueda
mesh_terms: D000581:Amidohydrolases / Q000378:metabolism*; D000818:Animals; D004789:Enzyme Activation; D057075:Enzyme Assays; D057809:HEK293 Cells; D006801:Humans; D007553:Isotope Labeling; D008168:Lung / Q000201:enzymology; D008297:Male; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D011994:Recombinant Proteins; D000581:Amidohydrolases; C575348:N-acylethanolamine-hydrolyzing acid amidase, rat
keywords: Anti-inflammatory agent; HEK293 cell; N-acylethanolamine-hydrolyzing acid amidase; NAAA; Palmitoylethanolamide; Radioisotope; Rat lung; Thin-layer chromatography