Assay of NAPE-PLD Activity

Filomena Fezza; Nicolina Mastrangelo; Mauro Maccarrone

doi:10.1007/978-1-4939-3539-0_13

Assay of NAPE-PLD Activity

Methods Mol Biol. 2016:1412:123-30. doi: 10.1007/978-1-4939-3539-0_13.

Authors

Filomena Fezza¹, Nicolina Mastrangelo², Mauro Maccarrone^{2

3}

Affiliations

¹ Department of Experimental Medicine and Surgery, Tor Vergata University of Rome, Rome, Italy. [email protected].
² Center of Integrated Research, Campus Bio-Medico University of Rome, Rome, Italy.
³ European Center for Brain Research/IRCCS Santa Lucia Foundation, Rome, Italy.

PMID: 27245898
DOI: 10.1007/978-1-4939-3539-0_13

Abstract

N-acyl-phosphatidylethanolamine (NAPE)-hydrolyzing phospholipase D (NAPE-PLD) is a prominent enzyme involved in the biosynthesis of fatty acid amides (FAAs), a family of bioactive lipids including anandamide (AEA) as the prototypical member. Here, we describe a NAPE-PLD assay based on radioactive substrates and product separation by thin-layer chromatography (TLC).

Keywords: Anandamide; Fatty acid amides; N-acyl-phosphatidylethanolamine-hydrolyzing phospholipase D; N-acyl-phosphatidylethanolamines; N-palmitoylethanolamine; Radioactive assay; Thin-layer chromatography.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Chromatography, Thin Layer
Enzyme Activation
Enzyme Assays*
Isotope Labeling
Phosphatidylethanolamines / metabolism*
Phospholipase D / metabolism*
Substrate Specificity

Substances

Phosphatidylethanolamines
Phospholipase D