HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 281, Issue 14, 9066-9075, April 7, 2006

This Article Full Text Full Text (PDF) All Versions of this Article: 281/14/9066    most recent M509721200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kaczocha, M. Articles by Deutsch, D. G. Search for Related Content PubMed PubMed Citation Articles by Kaczocha, M. Articles by Deutsch, D. G. Social Bookmarking                      What's this?

Anandamide Uptake Is Consistent with Rate-limited Diffusion and Is Regulated by the Degree of Its Hydrolysis by Fatty Acid Amide Hydrolase^*

Martin Kaczocha, Anita Hermann, Sherrye T. Glaser, Inge N. Bojesen^¶, and Dale G. Deutsch¹

From the Department of Biochemistry and Cell Biology, State University of New York at Stony Brook, Stony Brook, New York 11794-5215, the Center for Translational Neuroimaging, Brookhaven National Laboratory, Upton, New York 11973, and the ^¶Department of Medical Biochemistry and Genetics, University of Copenhagen, DK-2200 Copenhagen N, Denmark

The uptake of arachidonoyl ethanolamide (anandamide, AEA) in rat basophilic leukemia cells (RBL-2H3) has been proposed to occur via a saturable transporter that is blocked by specific inhibitors. Measuring uptake at 25 s, when fatty acid amide hydrolase (FAAH) does not appreciably affect uptake, AEA accumulated via a nonsaturable mechanism at 37 °C. Interestingly, saturation was observed when uptake was plotted using unbound AEA at 37 °C. Such apparent saturation can be explained by rate-limited delivery of AEA through an unstirred water layer surrounding the cells (1). In support of this, we observed kinetics consistent with rate-limited diffusion at 0 °C. Novel transport inhibitors have been synthesized that are either weak FAAH inhibitors or do not inhibit FAAH in vitro (e.g. UCM707, OMDM2, and AM1172). In the current study, none of these purported AEA transporter inhibitors affected uptake at 25 s. Longer incubation times illuminate downstream events that drive AEA uptake. Unlike the situation at 25 s, the efficacy of these inhibitors was unmasked at 5 min with appreciable inhibition of AEA accumulation correlating with partial inhibition of AEA hydrolysis. The uptake and hydrolysis profiles observed with UCM707, VDM11, OMDM2, and AM1172 mirrored two selective and potent FAAH inhibitors CAY10400 and URB597 (at low concentrations), indicating that weak inhibition of FAAH can have a pronounced effect upon AEA uptake. At 5 min, the putative transport inhibitors did not reduce AEA uptake in FAAH chemical knock-out cells. This strongly suggests that the target of UCM707, VDM11, OMDM2, and AM1172 is not a transporter at the plasma membrane but rather FAAH, or an uncharacterized intracellular component that delivers AEA to FAAH. This system is therefore unique among neuro/immune modulators because AEA, an uncharged hydrophobic molecule, diffuses into cells and partial inhibition of FAAH has a pronounced effect upon its uptake.

Received for publication, September 2, 2005 , and in revised form, January 11, 2006.

^* This work was supported by National Institutes of Health Grants DA9374 and DA16419. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed: Dept. of Biochemistry and Cell Biology, SUNY at Stony Brook, Stony Brook, NY 11794-5215. Tel.: 631-632-8595; Fax: 631-632-8575; E-mail: DDeutsch{at}notes.sunysb.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. L. Yates and E. L. Barker Inactivation and Biotransformation of the Endogenous Cannabinoids Anandamide and 2-Arachidonoylglycerol Mol. Pharmacol., July 1, 2009; 76(1): 11 - 17. [Abstract] [Full Text] [PDF]

				M. Kaczocha, S. T. Glaser, and D. G. Deutsch Identification of intracellular carriers for the endocannabinoid anandamide PNAS, April 14, 2009; 106(15): 6375 - 6380. [Abstract] [Full Text] [PDF]

				R. Russo, J. LoVerme, G. La Rana, T. R. Compton, J. Parrott, A. Duranti, A. Tontini, M. Mor, G. Tarzia, A. Calignano, et al. The Fatty Acid Amide Hydrolase Inhibitor URB597 (Cyclohexylcarbamic Acid 3'-Carbamoylbiphenyl-3-yl Ester) Reduces Neuropathic Pain after Oral Administration in Mice J. Pharmacol. Exp. Ther., July 1, 2007; 322(1): 236 - 242. [Abstract] [Full Text] [PDF]

				M. Bari, P. Spagnuolo, F. Fezza, S. Oddi, N. Pasquariello, A. Finazzi-Agro, and M. Maccarrone Effect of Lipid Rafts on Cb2 Receptor Signaling and 2-Arachidonoyl-Glycerol Metabolism in Human Immune Cells J. Immunol., October 15, 2006; 177(8): 4971 - 4980. [Abstract] [Full Text] [PDF]

				H. Wang, S. K. Dey, and M. Maccarrone Jekyll and Hyde: Two Faces of Cannabinoid Signaling in Male and Female Fertility Endocr. Rev., August 1, 2006; 27(5): 427 - 448. [Abstract] [Full Text] [PDF]