Crystal Structure of Fatty Acid Amide Hydrolase Bound to the Carbamate Inhibitor URB597: Discovery of a Deacylating Water Molecule and Insight into Enzyme Inactivation

JOURNAL OF MOLECULAR BIOLOGY

卷:400

Article

Mileni, Mauro¹  Kamtekar, Satwik²  Wood, David C.²  Benson, Timothy E.²  Cravatt, Benjamin F.³  Stevens, Raymond C.¹ 

[1] Scripps Res Inst, Dept Mol Biol, La Jolla, CA 92037 USA

[2] Pfizer Global Res & Dev, Chesterfield, MO 63017 USA

[3] Scripps Res Inst, Dept Physiol Chem, La Jolla, CA 92037 USA

关键词: fatty acid amide hydrolase (FAAH);    URB597;    crystal structure;    deacylating water;    catalytic mechanism;   

DOI  :  10.1016/j.jmb.2010.05.034

来源: Elsevier

【 摘 要 】

The endocannabinoid system regulates a wide range of physiological processes including pain, inflammation, and cognitive/emotional states. URB597 is one of the best characterized covalent inhibitors of the endocannabinoid-degrading enzyme fatty acid amide hydrolase (FAAH). Here, we report the structure of the FAAH URB597 complex at 2.3 angstrom resolution. The structure provides insights into mechanistic details of enzyme inactivation and experimental evidence of a previously uncharacterized active site water molecule that likely is involved in substrate deacylation. This water molecule is part of an extensive hydrogen-bonding network and is coordinated indirectly to residues lining the cytosolic port of the enzyme. In order to corroborate our hypothesis concerning the role of this water molecule in FAAH's catalytic mechanism, we determined the structure of FAAH conjugated to a urea-based inhibitor, PF-3845, to a higher resolution (2.4 angstrom) than previously reported. The higher-resolution structure confirms the presence of the water molecule in a virtually identical location in the active site. Examination of the structures of serine hydrolases that are non-homologous to FAAH, such as elastase, trypsin, or chymotrypsin, shows a similarly positioned hydrolytic water molecule and suggests a functional convergence between the amidase signature enzymes and serine proteases. (C) 2010 Elsevier Ltd. All rights reserved.

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