The objective of this proposal is to understand the catalytic, structural, and cell biological features of fatty acid amide hydrolase (FAAH), a mammalian membrane-bound enzyme responsible for the catabolism of the fatty acid amide family of endogenous signaling lipids. Representative fatty acid amides degraded by FAAH include the endocannabinoid anandamide and the sleep-inducing lipid oleamide. Fatty acid amides have been shown to induce a remarkable array of pharmacological effects in mammals, including sleep, analgesia, hypothermia, and learning and memory defects. The impressive bioactivity of fatty acid amides suggests that FAAH might serve as an attractive target for therapeutic efforts aimed at influencing pain, sleep, and memory systems. The elucidation of FAAH's catalytic, structural, and cellular features would provide a foundation for the design of FAAH-specific chemical inhibitors to be employed as agents for both the study of fatty acid amide-based physiological processes and the potential pharmaceutical treatment of pathologies associated with these systems. In this application, the molecular and cellular properties of FAAH will be examined using a multidisciplinary approach, employing biochemistry, molecular biology, immunochemistry, and synthetic chemistry techniques towards the goals of determining: 1) the catalytic mechanism and origins of substrate selectivity for FAAH-mediated amide hydrolysis, 2) the x-ray crystal structure of a FAAH-oleyl phosphonate inhibitor complex, 3) the domains of FAAH responsible for self-association and membrane binding, and 4) the cellular and subcellular localization of FAAH in mammalian tissue. These proposed studies should provide molecular tools for the chemical and genetic regulation of FAAH in vivo, allowing for a direct evaluation of the potential costs and benefits of therapeutic strategies that target the endocannabinioid system for the treatment of pain, sleep, and/or mood disorders.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Research Project (R01)
Project #
5R01DA013173-04
Application #
6693445
Study Section
Bio-Organic and Natural Products Chemistry Study Section (BNP)
Program Officer
Rapaka, Rao
Project Start
2001-02-01
Project End
2004-12-31
Budget Start
2004-01-01
Budget End
2004-12-31
Support Year
4
Fiscal Year
2004
Total Cost
$324,100
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
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Gulyas, A I; Cravatt, B F; Bracey, M H et al. (2004) Segregation of two endocannabinoid-hydrolyzing enzymes into pre- and postsynaptic compartments in the rat hippocampus, cerebellum and amygdala. Eur J Neurosci 20:441-58
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Cravatt, Benjamin F; Lichtman, Aron H (2003) Fatty acid amide hydrolase: an emerging therapeutic target in the endocannabinoid system. Curr Opin Chem Biol 7:469-75
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Leung, Donmienne; Hardouin, Christophe; Boger, Dale L et al. (2003) Discovering potent and selective reversible inhibitors of enzymes in complex proteomes. Nat Biotechnol 21:687-91
McKinney, Michele K; Cravatt, Benjamin F (2003) Evidence for distinct roles in catalysis for residues of the serine-serine-lysine catalytic triad of fatty acid amide hydrolase. J Biol Chem 278:37393-9
Lichtman, Aron H; Hawkins, E Gregory; Griffin, Graeme et al. (2002) Pharmacological activity of fatty acid amides is regulated, but not mediated, by fatty acid amide hydrolase in vivo. J Pharmacol Exp Ther 302:73-9
Sipe, Jack C; Chiang, Kyle; Gerber, Alexandra L et al. (2002) A missense mutation in human fatty acid amide hydrolase associated with problem drug use. Proc Natl Acad Sci U S A 99:8394-9
Waleh, N S; Cravatt, B F; Apte-Deshpande, A et al. (2002) Transcriptional regulation of the mouse fatty acid amide hydrolase gene. Gene 291:203-10

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