title: Hydrolysis of ibuprofenoyl-CoA and other 2-APA-CoA esters by human acyl-CoA thioesterases-1 and -2 and their possible role in the chiral inversion of profens.
abstract: Ibuprofen and related 2-arylpropanoic acid (2-APA) drugs are often given as a racemic mixture and the R-enantiomers undergo activation in vivo by metabolic chiral inversion. The chiral inversion pathway consists of conversion of the drug to the coenzyme A ester (by an acyl-CoA synthetase) followed by chiral inversion by α-methylacyl-CoA racemase (AMACR; P504S). The enzymes responsible for hydrolysis of the product S-2-APA-CoA ester to the active S-2-APA drug have not been identified. In this study, conversion of a variety of 2-APA-CoA esters by human acyl-CoA thioesterase-1 and -2 (ACOT-1 and -2) was investigated. Human recombinant ACOT-1 and -2 (ACOT-1 and -2) were both able to efficiently hydrolyse a variety of 2-APA-CoA substrates. Studies with the model substrates R- and S-2-methylmyristoyl-CoA showed that both enzymes were able to efficiently hydrolyse both of the epimeric substrates with (2R)- and (2S)- methyl groups. ACOT-1 is located in the cytosol and is able to hydrolyse 2-APA-CoA esters exported from the mitochondria and peroxisomes for inhibition of cyclo-oxygenase-1 and -2 in the endoplasmic reticulum. It is a prime candidate to be the enzyme responsible for the pharmacological action of chiral inverted drugs. ACOT-2 activity may be important in 2-APA toxicity effects and for the regulation of mitochondrial free coenzyme A levels. These results support the idea that 2-APA drugs undergo chiral inversion via a common pathway.
authors.affiliation: Medicinal Chemistry, Department of Pharmacy and Pharmacology, University of Bath, Claverton Down, Bath BA2 7AY, UK.
authors.forename: Xiao
authors.initials: X
authors.lastname: Qu
authors.forename: Amanda
authors.initials: A
authors.lastname: Allan
authors.forename: Grace
authors.initials: G
authors.lastname: Chui
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Hutchings
authors.forename: Ping
authors.initials: P
authors.lastname: Jiao
authors.forename: Lawrence
authors.initials: L
authors.lastname: Johnson
authors.forename: Wai Y
authors.initials: WY
authors.lastname: Leung
authors.forename: Portia K
authors.initials: PK
authors.lastname: Li
authors.forename: Georgina R
authors.initials: GR
authors.lastname: Steel
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Thompson
authors.forename: Michael D
authors.initials: MD
authors.lastname: Threadgill
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Woodman
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Lloyd
mesh_terms: D000214:Acyl Coenzyme A / Q000737:chemistry* / Q000378:metabolism; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics; D004952:Esters; D006801:Humans; D006868:Hydrolysis; D007052:Ibuprofen / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D015394:Molecular Structure; D010170:Palmitoyl-CoA Hydrolase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D013237:Stereoisomerism; D013379:Substrate Specificity; D013869:Thiolester Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D000214:Acyl Coenzyme A; D004952:Esters; C070652:ibuprofenyl-coenzyme A; D013869:Thiolester Hydrolases; C413373:ACOT2 protein, human; D010170:Palmitoyl-CoA Hydrolase; D007052:Ibuprofen
keywords: 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid; 5,5′-dithiobis(2-nitrobenzoic acid); ACOT; Acyl-CoA thioesterase (ACOT); BSA; Branched-chain fatty acids; DTNB; HEPES; Ibuprofen; NSAID; NSAIDs; P504S); PMSF; SDS-PAGE; acyl-CoA thioesterase; bovine serum albumin; non-steroidal anti-inflammatory drugs; phenylmethylsulfonyl fluoride; sodium dodecyl sulfate polyacrylamide gel electrophoresis; α-Methylacyl-CoA racemase (AMACR