title: A catalytically silent FAAH-1 variant drives anandamide transport in neurons.
abstract: The endocannabinoid anandamide is removed from the synaptic space by a selective transport system, expressed in neurons and astrocytes, that remains molecularly uncharacterized. Here we describe a partly cytosolic variant of the intracellular anandamide-degrading enzyme fatty acid amide hydrolase-1 (FAAH-1), termed FAAH-like anandamide transporter (FLAT), that lacked amidase activity but bound anandamide with low micromolar affinity and facilitated its translocation into cells. Known anandamide transport inhibitors, such as AM404 and OMDM-1, blocked these effects. We also identified a competitive antagonist of the interaction of anandamide with FLAT, the phthalazine derivative ARN272, that prevented anandamide internalization in vitro, interrupted anandamide deactivation in vivo and exerted profound analgesic effects in rodent models of nociceptive and inflammatory pain, which were mediated by CB(1) cannabinoid receptors. The results identify FLAT as a critical molecular component of anandamide transport in neural cells and a potential target for therapeutic drugs.
authors.affiliation: Department of Pharmacology, University of California, Irvine, Irvine, California, USA.
authors.forename: Jin
authors.initials: J
authors.lastname: Fu
authors.forename: Giovanni
authors.initials: G
authors.lastname: Bottegoni
authors.forename: Oscar
authors.initials: O
authors.lastname: Sasso
authors.forename: Rosalia
authors.initials: R
authors.lastname: Bertorelli
authors.forename: Walter
authors.initials: W
authors.lastname: Rocchia
authors.forename: Matteo
authors.initials: M
authors.lastname: Masetti
authors.forename: Ana
authors.initials: A
authors.lastname: Guijarro
authors.forename: Alessio
authors.initials: A
authors.lastname: Lodola
authors.forename: Andrea
authors.initials: A
authors.lastname: Armirotti
authors.forename: Gianpiero
authors.initials: G
authors.lastname: Garau
authors.forename: Tiziano
authors.initials: T
authors.lastname: Bandiera
authors.forename: Angelo
authors.initials: A
authors.lastname: Reggiani
authors.forename: Marco
authors.initials: M
authors.lastname: Mor
authors.forename: Andrea
authors.initials: A
authors.lastname: Cavalli
authors.forename: Daniele
authors.initials: D
authors.lastname: Piomelli
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism* / Q000494:pharmacology; D001593:Benzyl Compounds / Q000494:pharmacology; D063388:Endocannabinoids; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D053284:Polyunsaturated Alkamides / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects / Q000502:physiology; D051381:Rats; D000099252:Fatty Acid Amide Hydrolases
chemical_list: D000581:Amidohydrolases; D001095:Arachidonic Acids; D001593:Benzyl Compounds; D063388:Endocannabinoids; D053284:Polyunsaturated Alkamides; D000099252:Fatty Acid Amide Hydrolases; C474344:OMDM-1 cpd; C078814:anandamide; C107594:N-(4-hydroxyphenyl)arachidonylamide
references.citation: Proc Natl Acad Sci U S A. 2001 Jul 31;98(16):9371-6
references.pmid: 11470906
references.citation: Chem Rev. 2007 Aug;107(8):3687-714
references.pmid: 17683161
references.citation: Chem Biol. 2009 Jun 26;16(6):624-32
references.pmid: 19481477
references.citation: Nat Rev Neurosci. 2007 Oct;8(10):743-54
references.pmid: 17882252
references.citation: Biochemistry. 2004 Jun 29;43(25):8184-90
references.pmid: 15209515
references.citation: J Pharmacol Exp Ther. 2007 Jul;322(1):236-42
references.pmid: 17412883
references.citation: Science. 1992 Dec 18;258(5090):1946-9
references.pmid: 1470919
references.citation: Trends Biochem Sci. 2010 Nov;35(11):601-8
references.pmid: 20570522
references.citation: J Lipid Res. 2003 Sep;44(9):1790-4
references.pmid: 12837852
references.citation: Neuroreport. 2000 Apr 27;11(6):1231-5
references.pmid: 10817598
references.citation: Int J Biol Macromol. 2008 Jan 1;42(1):62-7
references.pmid: 17996290
references.citation: J Pharmacol Exp Ther. 2004 Nov;311(2):441-8
references.pmid: 15229230
references.citation: J Biol Chem. 2006 Dec 1;281(48):36569-78
references.pmid: 17015445
references.citation: Biochem Pharmacol. 2003 May 1;65(9):1473-81
references.pmid: 12732359
references.citation: J Mol Neurosci. 2007 Sep;33(1):18-24
references.pmid: 17901541
references.citation: Science. 1997 Aug 22;277(5329):1094-7
references.pmid: 9262477
references.citation: Eur J Neurosci. 2004 Jul;20(2):441-58
references.pmid: 15233753
references.citation: Br J Pharmacol. 2008 Nov;155(5):775-82
references.pmid: 18660824
references.citation: Biochemistry. 1999 Oct 26;38(43):14125-30
references.pmid: 10571985
references.citation: J Neurochem. 1997 Aug;69(2):631-8
references.pmid: 9231721
references.citation: Cell Metab. 2008 Oct;8(4):281-288
references.pmid: 18840358
references.citation: Eur J Biochem. 2001 Apr;268(7):1982-9
references.pmid: 11277920
references.citation: Annu Rev Biochem. 2005;74:411-32
references.pmid: 15952893
references.citation: Biochem J. 2004 May 15;380(Pt 1):265-72
references.pmid: 14969584
references.citation: Br J Pharmacol. 2001 Apr;132(8):1743-54
references.pmid: 11309246
references.citation: Science. 2002 Nov 29;298(5599):1793-6
references.pmid: 12459591
references.citation: Biochim Biophys Acta. 1999 Nov 23;1441(2-3):106-16
references.pmid: 10570239
references.citation: Proc Natl Acad Sci U S A. 2003 Apr 1;100(7):4269-74
references.pmid: 12655057
references.citation: Nat Rev Drug Discov. 2008 May;7(5):438-55
references.pmid: 18446159
references.citation: Proc Natl Acad Sci U S A. 1999 May 11;96(10):5802-7
references.pmid: 10318965
references.citation: J Comput Chem. 2008 Jan 15;29(1):24-37
references.pmid: 17910006
references.citation: Nat Med. 2003 Jan;9(1):76-81
references.pmid: 12461523
references.citation: Br J Pharmacol. 2003 Nov;140(5):802-8
references.pmid: 12970089
references.citation: Prog Lipid Res. 2009 Mar;48(2):101-15
references.pmid: 19280719
references.citation: J Pharmacol Exp Ther. 2006 Jun;317(3):1365-71
references.pmid: 16510698
references.citation: Mol Pharmacol. 2009 Jul;76(1):11-7
references.pmid: 19389920
references.citation: Chem Biol. 2005 Nov;12(11):1179-87
references.pmid: 16298297
references.citation: Nat Rev Mol Cell Biol. 2005 May;6(5):386-98
references.pmid: 15956978
references.citation: Science. 1995 May 12;268(5212):866-9
references.pmid: 7754369