Literature summary for 3.5.1.99 extracted from

Patricelli, M.P.; Cravatt, B.F.
Clarifying the catalytic roles of conserved residues in the amidase signature family (2000), J. Biol. Chem., 275, 19177-19184.

Cloned(Commentary)

Cloned (Comment)	Organism
FAAH mutants are expressed in the Escherichia coli strain BL21. All Escherichia coli expression constructs contained a deletion of the N-terminal transmembrane domain of FAAH. Deletion of this region facilitates its purification from Escherichia coli but has no effect on the enzymatic activity of FAAH. Some mutants are expressed mainly as inclusion body in Escherichia coli, preventing a detailed analysis of their catalytic function in this system. The expression of FAAH mutants in COS-7 cells provides a system where the majority of these variants can be directly compared	Rattus norvegicus

Cloned (Comment)

Organism

FAAH mutants are expressed in the Escherichia coli strain BL21. All Escherichia coli expression constructs contained a deletion of the N-terminal transmembrane domain of FAAH. Deletion of this region facilitates its purification from Escherichia coli but has no effect on the enzymatic activity of FAAH. Some mutants are expressed mainly as inclusion body in Escherichia coli, preventing a detailed analysis of their catalytic function in this system. The expression of FAAH mutants in COS-7 cells provides a system where the majority of these variants can be directly compared

Rattus norvegicus

Protein Variants

Protein Variants	Comment	Organism
D167A	mutant enzyme shows 48% of wild-type activity with oleamide as substrate	Rattus norvegicus
D237E	mutant enzyme shows 12% of wild-type activity with oleamide as substrate	Rattus norvegicus
D237N	mutant enzyme shows 2.3% of wild-type activity with oleamide as substrate	Rattus norvegicus
E143Q	mutant enzyme shows 60% of wild-type activity with oleamide as substrate	Rattus norvegicus
K142A	mutation decreases the amidase activity of FAAH greater than 100fold without detectably impacting the structural integrity of the enzyme, mutant enzyme shows 3% of the oleoyl ester hydrolysis compared to wild-type enzyme	Rattus norvegicus
K142A/R243A	no hydrolysis of oleoyl methyl ester	Rattus norvegicus
K255A	mutant enzyme shows 17% of wild-type activity with oleamide as substrate	Rattus norvegicus
N206A	mutant enzyme shows 11% of wild-type activity with oleamide as substrate	Rattus norvegicus
R243A	mutation decreases the amidase activity of FAAH greater than 100fold without detectably impacting the structural integrity of the enzyme, mutant enzyme shows 24% of the oleoyl ester hydrolysis compared to wild-type enzyme	Rattus norvegicus
S217A	mutation decreases the amidase activity of FAAH greater than 100fold without detectably impacting the structural integrity of the enzyme, no hydrolysis of oleoyl methyl ester	Rattus norvegicus
S218A	mutation decreases the amidase activity of FAAH greater than 100fold without detectably impacting the structural integrity of the enzyme, mutant enzyme shows 3% of the oleoyl ester hydrolysis compared to wild-type enzyme	Rattus norvegicus
S241A	mutation decreases the amidase activity of FAAH greater than 100fold without detectably impacting the structural integrity of the enzyme, no hydrolysis of oleoyl methyl ester	Rattus norvegicus
T257A	mutant enzyme shows 65% of wild-type activity with oleamide as substrate	Rattus norvegicus

Localization

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

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
anandamide + H2O	Rattus norvegicus	the enzyme is responsible for the hydrolysis of a number of neuromodulatory fatty acid amides, including the endogenous cannabinoid anandamide and the sleep-inducing lipid oleamide	ethanolamine + arachidonic acid	-	?
oleamide + H2O	Rattus norvegicus	the enzyme is responsible for the hydrolysis of a number of neuromodulatory fatty acid amides, including the endogenous cannabinoid anandamide and the sleep-inducing lipid oleamide	oleic acid + NH3	-	?

Organism

Organism	UniProt	Comment	Textmining
Rattus norvegicus	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
all Escherichia coli expression constructs contained a deletion of the N-terminal transmembrane domain of FAAH. Deletion of this region facilitates its purification from Escherichia coli but had no effect on the enzymatic activity of FAAH	Rattus norvegicus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
anandamide + H2O	the enzyme is responsible for the hydrolysis of a number of neuromodulatory fatty acid amides, including the endogenous cannabinoid anandamide and the sleep-inducing lipid oleamide	Rattus norvegicus	ethanolamine + arachidonic acid	-	?
oleamide + H2O	-	Rattus norvegicus	oleic acid + NH3	-	?
oleamide + H2O	the enzyme is responsible for the hydrolysis of a number of neuromodulatory fatty acid amides, including the endogenous cannabinoid anandamide and the sleep-inducing lipid oleamide	Rattus norvegicus	oleic acid + NH3	-	?
oleoyl methyl ester + H2O	-	Rattus norvegicus	oleic acid + methanol	-	?

Synonyms

Synonyms	Comment	Organism
FAAH	-	Rattus norvegicus

pH Range

pH Minimum	pH Maximum	Comment	Organism
additional information	-	pH rate profiles of FAAH mutants	Rattus norvegicus

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
5	-	not stable below	Rattus norvegicus