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. 2023:2576:261-274.
doi: 10.1007/978-1-0716-2728-0_22.

Assay of NAAA Activity

Kazuhito Tsuboi  1 , Natsuo Ueda  2
Affiliations

Assay of NAAA Activity

Kazuhito Tsuboi et al. Methods Mol Biol. 2023.

Abstract

N-acylethanolamine-hydrolyzing acid amidase (NAAA) is a lysosomal hydrolase degrading various N-acylethanolamines at acidic pH. NAAA prefers anti-inflammatory and analgesic palmitoylethanolamide to other N-acylethanolamines as a substrate, and its specific inhibitors are shown to exert anti-inflammatory and analgesic actions in animal models. Therefore, these inhibitors are expected as a new class of therapeutic agents. Here, we introduce an NAAA assay system, using [14C]palmitoylethanolamide and thin-layer chromatography. The preparation of NAAA enzyme from native and recombinant sources as well as the chemical synthesis of N-[1'-14C]palmitoyl-ethanolamine is also described.

Keywords: Anti-inflammatory agent; HEK293 cell; N-acylethanolamine acid amidase; NAAA; Palmitoylethanolamide; Radioisotope; Rat lung; Thin-layer chromatography.

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References

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