Literature summary for 3.5.1.99 extracted from

Mileni, M.; Kamtekar, S.; Wood, D.C.; Benson, T.E.; Cravatt, B.F.; Stevens, R.C.
Crystal structure of fatty acid amide hydrolase bound to the carbamate inhibitor URB597: discovery of a deacylating water molecule and insight into enzyme inactivation (2010), J. Mol. Biol., 400, 743-754.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of mutant L192F/F194Y/A377T/S435N/I491V/V495M in Escherichia coli strain BL21	Rattus norvegicus

Crystallization (Commentary)

Crystallization (Comment)	Organism
recombinant fatty acid amide hydrolase mutant L192F/F194Y/A377T/S435N/I491V/V495M bound to carbamate inhibitor URB597 or inhibitor PF-3845, 25-30 mg/ml protein in 10 mM HEPES, pH 7.0, 500 mM NaCl, 2 mM dithiothreitol, and 0.08% n-undecyl-beta-D-maltoside or 0.1% n-decyl-beta-D-maltoside for inhibitor URB597-enzyme or PF-3845-enzyme conjugate, respectively, supplementation of the FAAHURB597 proteinsample with 1.6% benzyldimethyl(2-dodecyloxyethyl)-ammonium chloride and mixed at a 1:1 proportion with a crystallization buffer containing 30% PEG 400, 100 mM TrisHCl pH 7.5, and 100 mM MgCl2, supplementation of the FAAHPF-3845 protein sample with 1.6% benzyl-dimethyl-dodecyl ammonium bromide and mixed at a 1:1 ratio with a crystallization buffer containing 30% PEG 400, 100 mM 2-(N-morpholino)ethanesulfonate/NaOH, pH 5.5, and 400 mM LiCl, sitting drop vapor diffusion at 14°C, X-ray diffraction structure determination and analysis at 2.3 A resolution	Rattus norvegicus

Crystallization (Comment)

Organism

recombinant fatty acid amide hydrolase mutant L192F/F194Y/A377T/S435N/I491V/V495M bound to carbamate inhibitor URB597 or inhibitor PF-3845, 25-30 mg/ml protein in 10 mM HEPES, pH 7.0, 500 mM NaCl, 2 mM dithiothreitol, and 0.08% n-undecyl-beta-D-maltoside or 0.1% n-decyl-beta-D-maltoside for inhibitor URB597-enzyme or PF-3845-enzyme conjugate, respectively, supplementation of the FAAHURB597 proteinsample with 1.6% benzyldimethyl(2-dodecyloxyethyl)-ammonium chloride and mixed at a 1:1 proportion with a crystallization buffer containing 30% PEG 400, 100 mM TrisHCl pH 7.5, and 100 mM MgCl2, supplementation of the FAAHPF-3845 protein sample with 1.6% benzyl-dimethyl-dodecyl ammonium bromide and mixed at a 1:1 ratio with a crystallization buffer containing 30% PEG 400, 100 mM 2-(N-morpholino)ethanesulfonate/NaOH, pH 5.5, and 400 mM LiCl, sitting drop vapor diffusion at 14°C, X-ray diffraction structure determination and analysis at 2.3 A resolution

Rattus norvegicus

Protein Variants

Protein Variants	Comment	Organism
L192F/F194Y/A377T/S435N/I491V/V495M	generation of a humanized version of the rat FAAH by replacment of six amino acids in the active site of the rat FAAH protein in order to recreate the binding profile of the human enzyme	Rattus norvegicus

Inhibitors

Inhibitors	Comment	Organism
PF-3845	crystal structure analysis of enzyme-inhibitor complex. The inhibitor interacts with the catalytic triad and oxyanion hole residues	Rattus norvegicus
URB524	-	Rattus norvegicus
URB597	a carbamate inhibitor that displays excellent selectivity for FAAH in the nervous system, although the inhibitor does inactivate additional peripheral hydrolases, enzyme-inhibitor binding structure, overview. The inhibitor interacts with the catalytic triad and oxyanion hole residues	Rattus norvegicus

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
membrane	-	Rattus norvegicus	16020	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
anandamide + H2O	Rattus norvegicus	i.e. arachidonoyl ethanolamide	arachidonic acid + ethanolamine	-	?
oleamide + H2O	Rattus norvegicus	i.e. cis-9-octadecenamide	oleic acid + NH3	-	?

Organism

Organism	UniProt	Comment	Textmining
Rattus norvegicus	P97612	recombinant mutant L192F/F194Y/A377T/S435N/I491V/V495M, a humanized version of the rat FAAH	-

Reaction

Reaction	Comment	Organism	Reaction ID
anandamide + H2O = arachidonic acid + ethanolamine	structure-function relationship and catalytic mechanism, overview	Rattus norvegicus	a
oleamide + H2O = oleic acid + NH3	structure-function relationship and catalytic mechanism, overview	Rattus norvegicus	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
anandamide + H2O	i.e. arachidonoyl ethanolamide	Rattus norvegicus	arachidonic acid + ethanolamine	-	?
oleamide + H2O	i.e. cis-9-octadecenamide	Rattus norvegicus	oleic acid + NH3	-	?

Synonyms

Synonyms	Comment	Organism
FAAH	-	Rattus norvegicus

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor	Structure
0.0000046	-	pH and temperature not specified in the publication	Rattus norvegicus	URB597
0.000063	-	pH and temperature not specified in the publication	Rattus norvegicus	URB524

General Information

General Information	Comment	Organism
evolution	FAAH is a member of the amidase signature family found in bacteria, archaea and eukaryotes	Rattus norvegicus
physiological function	fatty acid amide hydrolase degrades lipid signaling molecules such as the endogenous cannabinoids anandamide, i.e. N-arachidonyl ethanolamine, and the sleep-inducing substance oleamide, i.e. cis-9-octadecenamide	Rattus norvegicus