Anandamide hydrolysis by human cells in culture and brain

Mauro Maccarrone, Marcelis Van Der Stelt, Antonello Rossi, Gerrit A. Veldink, J. F G Vliegenthart, Alessandro Finazzi Agrò

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Abstract

Anandamide (arachidonylethanolamide; AnNH) has important neuromodulatory and immunomodulatory activities. This lipid is rapidly taken up and hydrolyzed to arachidonate and ethanolamine in many organisms. As yet, AnNH inactivation has not been studied in humans. Here, a human brain fatty-acid amide hydrolase (FAAH) has been characterized as a single protein of 67 kDa with a pI of 7.6, showing apparent K(m) and V(max) values for AnNH of 2.0 ± 0.2 μM and 800 ± 75 pmol·min-1·mg of protein-1, respectively. The optimum pH and temperature for AnNH hydrolysis were 9.0 and 37 °C, respectively, and the activation energy of the reaction was 43.5 ± 4.5 kJ·mol-1. Hydro(pero)xides derived from AnNH or its linoleoyl analogues by lipoxygenase action were competitive inhibitors of human brain FAAH, with apparent K(i) values in the low micromolar range. One of these compounds, linoleoylethanolamide is the first natural inhibitor (K(i) = 9.0 ± 0.9 μM) of FAAH as yet discovered. An FAAH activity sharing several biochemical properties with the human brain enzyme was demonstrated in human neuroblastoma CHP100 and lymphoma U937 cells. Both cell lines have a high affinity transporter for AnNH, which had apparent K(m) and V(max) values for AnNH of 0.20 ± 0.02 μM and 30 ± 3 pmol·min-1·mg of protein-1 (CHP100 cells) and 0.13 ± 0.01 μM and 140 ± 15 pmol·min-1·mg of protein-1 (U937 cells), respectively. The AnNH carrier of both cell lines was activated up to 170% of the control by nitric oxide.

Original languageEnglish
Pages (from-to)32332-32339
Number of pages8
JournalJournal of Biological Chemistry
Volume273
Issue number48
DOIs
Publication statusPublished - Nov 27 1999

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ASJC Scopus subject areas

  • Biochemistry

Cite this

Maccarrone, M., Van Der Stelt, M., Rossi, A., Veldink, G. A., Vliegenthart, J. F. G., & Agrò, A. F. (1999). Anandamide hydrolysis by human cells in culture and brain. Journal of Biological Chemistry, 273(48), 32332-32339. https://doi.org/10.1074/jbc.273.48.32332