Molecular Characterization of a Phospholipase D Generating Anandamide and Its Congeners*

By Y. Okamoto, J. Morishita, K. Tsuboi, T. Tonai, N. Ueda. Journal of Biological Chemistry, 2004

811 citations recorded by the source corpus.

Anandamide (N-arachidonoylethanolamine) is known to be an endogenous ligand of cannabinoid and vanilloid receptors. Its congeners (collectively referred to as N-acylethanolamines) also show a variety of biological activities. These compounds are principally formed from their corresponding N-acyl-phosphatidylethanolamines by a phosphodiesterase of the phospholipase D-type in animal tissues. We purified the enzyme from rat heart, and by the use of the sequences of its internal peptides cloned its complementary DNAs from mouse, rat, and human. The deduced amino acid sequences were composed of 393–396 residues, and showed that the enzyme has no homology with the known phospholipase D enzymes but is classified as a member of the zinc metallohydrolase family of the β-lactamase fold. As was overexpressed in COS-7 cells, the recombinant enzyme generated anandamide and other N-acylethanolamines from their corresponding N-acyl-phosphatidylethanolamines at comparable rates. In contrast, the enzyme was inactive with phosphatidylcholine and phosphatidylethanolamine. Assays of the enzyme activity and the messenger RNA and protein levels revealed its wide distribution in murine organs with higher contents in the brain, kidney, and testis. These results confirm that a specific phospholipase D is responsible for the generation of N-acylethanolamines including anandamide, strongly suggesting the physiological importance of lipid molecules of this class.

Subfields: Pathology and Forensic Medicine, Pharmacology, Toxicology

View Paper: https://www.semanticscholar.org/paper/1b01cf13abb2d2c89db62b729d7c432c3c4f6c84

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Pathology and Forensic Medicine

1. V. Di Marzo

In Pharmacology

1. V. Di Marzo
2. George Kunos
3. P. Pacher
4. S. Batkai
5. Hui-Chen Lu
6. L. Petrocellis
7. K. Mackie
8. M. Bifulco

In Toxicology

1. George Kunos
2. V. Di Marzo

Citations in the corpus

• The endocannabinoid system and its therapeutic exploitation (2004, positive): “but remained uncharacterized until only a few months ago, is N -acylphosphatidylethanolamine-selective phospholipase D (NAPE-PLD) ( Box 1”
• Endocannabinoid control of food intake and energy balance (2005, background): “Phospholipid-dependent pathways for endocannabinoid biosynthesis were discovered, leading to the cloning of the enzymes that catalyze the formation of anandamide and 2-AG from their direct precursors: the N -acylphosphatidylethanolamine-selective phospholipase D and the sn -1–selective diacylglyce”
• The Endocannabinoid System as an Emerging Target of Pharmacotherapy (2006, background): “Although a specific transacylase for the latter reaction has not yet been identified, a NAPE-specific PLD has recently been cloned (Okamoto et al., 2004).”
• Distribution of cannabinoid receptors in the central and peripheral nervous system. (2005, neutral): “phospholipase D (Okamoto et al. 2004) remain to be done.”
• An introduction to the endogenous cannabinoid system (2015, background): “A mammalian calcium-stimulated NAPE-PLD has been cloned and characterized (51).”