• Methods Mol. Biol. · Jan 2016

    Assay of NAAA Activity.

    • Kazuhito Tsuboi and Natsuo Ueda.
    • Department of Biochemistry, Kagawa University School of Medicine, 1750-1 Ikenobe, Miki, Kagawa, 761-0793, Japan.
    • Methods Mol. Biol. 2016 Jan 1; 1412: 137-48.

    AbstractN-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.

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