Online citations, reference lists, and bibliographies.
← Back to Search

Molecular Cloning And Expression Of A 2-arylpropionyl-coenzyme A Epimerase: A Key Enzyme In The Inversion Metabolism Of Ibuprofen.

C. Reichel, R. Brugger, H. Bang, G. Geisslinger, K. Brune
Published 1997 · Biology, Medicine

Cite This
Download PDF
Analyze on Scholarcy
The 2-arylpropionic acid derivatives, including ibuprofen, are the most widely used anti-inflammatory analgesic cyclooxygenase inhibitors. The (-)-R-enantiomer, which is inactive in terms of cyclooxygenase inhibition, is epimerized in vivo via the 2-arylpropionyl-coenzyme A (CoA) epimerase to the cyclooxygenase-inhibiting (+)-S-enantiomer. The molecular biology of the epimerization pathway is largely unknown. To clarify this mechanism, the sequence of the 2-arylpropionyl-CoA epimerase was identified, and the enzyme cloned and expressed. A cDNA clone encoding the 2-arylpropionyl-CoA epimerase was isolated from a rat liver cDNA library. The nucleotide and the deduced amino acid sequence of this enzyme was determined. Significant amino acid sequence similarity was found between the rat epimerase and carnitine dehydratases from Caenorhabditis elegans (41%) and Escherichia coli (27%). A bacterial expression system (E. coli strain M15[pREP4]) was used to express the epimerase protein, representing up to 20-30% of the total cellular E. coli protein. The expression of the epimerase was confirmed with Western blots using specific anti-epimerase antibodies and by measuring the rate of inversion of (R)-ibuprofenoyl-CoA. Northern blot analysis revealed a prominent 1.9-kb mRNA transcript in different rat tissues. In addition to its obvious importance in drug metabolism, the homology of the epimerase with carnitine dehydratases from several species suggests that this protein, which up to now has only been characterized as having a role in drug transformation, has a function in lipid metabolism.
This paper references
A Laboratory manual
M. Ashburner (1989)
Is the formation of R-ibuprofenyl-adenylate the first stereoselective step of chiral inversion?
S. Menzel (1994)
Microbial metabolism of 2-arylpropionic acids: chiral inversion of ibuprofen and 2-phenylpropionic acid.
A. J. Hutt (1993)
Structure and regulation of rat long-chain acyl-CoA synthetase.
H. Suzuki (1990)
Stereoselective arylpropionyl-CoA thioester formation in vitro.
M. Knadler (1990)
Antibodies: A Laboratory Manual
E. Harlow (1988)
Chiral inversion of 2-arylpropionic acid non-steroidal anti-inflammatory drugs--II. Racemization and hydrolysis of (R)- and (S)-ibuprofen-CoA thioesters.
R. Knihinicki (1991)
Mechanistic studies of the metabolic chiral inversion of (R)-ibuprofen in humans.
T. Baillie (1989)
Brownlee . 3 9 non - coding region sequences in eukaryotic messenger RNA
N. F. Proudfoot (1976)
Ibuprofen stereoisomer hepatic clearance and distribution in normal and fatty in situ perfused rat liver.
J. Cox (1985)
Metabolic inversion of (R)-ibuprofen. Formation of ibuprofenyl-coenzyme A.
T. Tracy (1993)
Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
P. Chomczyński (1987)
evidence for metabolic inversion of the (2)-isomer
S. S. Adams (1976)
Molecular characterization of the cai operon necessary for carnitine metabolism in Escherichia coii
K. Eichler (1994)
The dynamic role of palmitoylation in signal transduction.
G. Milligan (1995)
influence of protein binding
S. D. Hall (1992)
Stereoselective metabolism of 2-phenylpropionic acid in rat. I. In vitro studies on the stereoselective isomerization and glucuronidation of 2-phenylpropionic acid.
Y. Nakamura (1987)
Cyclooxygenase inhibition and the spinal release of prostaglandin E2 and amino acids evoked by paw formalin injection: a microdialysis study in unanesthetized rats
A. Malmberg (1995)
The metabolic chiral inversion of 2-phenylpropionic acid in rat, mouse and rabbit.
S. Fournel (1986)
TIS10, a phorbol ester tumor promoter-inducible mRNA from Swiss 3T3 cells, encodes a novel prostaglandin synthase/cyclooxygenase homologue.
D. Kujubu (1991)
Molecular Cloning: A Laboratory Manual
J. Sambrook (1983)
Pulmonary inversion of 2-arylpropionic acids: influence of protein binding.
S. Hall (1992)
A simple method for displaying the hydropathic character of a protein.
J. Kyte (1982)
The role of coenzyme A in the biotransformation of 2-arylpropionic acids.
S. Hall (1994)
Consensus sequences as substrate specificity determinants for protein kinases and protein phosphatases.
P. Kennelly (1991)
Studies on the metabolism and chiral inversion of ibuprofen in isolated rat hepatocytes.
S. M. Sanins (1990)
2.2 Mb of contiguous nucleotide sequence from chromosome III of C. elegans
R. Wilson (1994)
Metabolic inversion of (R)-ibuprofen. Epimerization and hydrolysis of ibuprofenyl-coenzyme A.
T. Tracy (1992)
Isolation and characterization of rat liver microsomal R-ibuprofenoyl-CoA synthetase.
R. Brugger (1996)
Isolation and characterization of rat liver microsomal Ribuprofenyl-CoA synthetase
R. Brugger (1996)
Metabolic stereoisomeric inversion of ibuprofen in mammals.
C. Chen (1991)
3′ Non-coding region sequences in eukaryotic messenger RNA
N. Proudfoot (1976)
Synthesis of L-carnitine by microorganisms and isolated enzymes.
H. Jung (1993)
Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs.
M. Kozak (1984)
Metabolic chiral inversion of 2-arylpropionates in rat H4IIE and human Hep G2 hepatoma cells. Relationship to in vivo metabolism.
S. Menzel-Soglowek (1992)
Pharmacological differences between the optical isomers of ibuprofen: evidence for metabolic inversion of the (—)‐isomer
S. S. Adams (1976)
Common Principles of Protein Translocation Across Membranes
G. Schatz (1996)
Stereoselective highperformance liquid chromatographic determination of ketoprofen, ibuprofen and fenoprofen in plasma using a chiral a1-acid glycoprotein
S. Menzel-Soglowek (1990)
Single step method of RNA extraction by guanidinium thiocyanate-phenol-chloroform
P. Chomczynski (1987)
Differential inhibition of prostaglandin endoperoxide synthase (cyclooxygenase) isozymes by aspirin and other non-steroidal anti-inflammatory drugs.
E. Meade (1993)
The metabolic chiral inversion and dispositional enantioselectivity of the 2-arylpropionic acids and their biological consequences.
J. Caldwell (1988)
Cloning and functional expression of a novel long-chain acyl-CoA synthetase expressed in brain.
T. Fujino (1992)
2-Arylpropionyl-CoA epimerase: partial peptide sequences and tissue localization.
C. Reichel (1995)
Stereoselective high-performance liquid chromatographic determination of ketoprofen, ibuprofen and fenoprofen in plasma using a chiral alpha 1-acid glycoprotein column.
S. Menzel-Soglowek (1990)
Purification and characterization of novel "2-arylpropionyl-CoA epimerases" from rat liver cytosol and mitochondria.
W. R. Shieh (1993)

This paper is referenced by
Računalne simulacije kompleksa derivata ibuprofena i naproksena s acil-CoA tioesterazom I (TesA) iz bakterije Pseudomonas aeruginosa
Krunoslav Ilić (2015)
Stereoselective Pharmacokinetics and Chiral Inversion of Ibuprofen in Adjuvant-induced Arthritic Rats
Hiroyuki Ikuta (2017)
Impaired intrinsic chiral inversion activity of ibuprofen in rats with adjuvant-induced arthritis
S. Uno (2008)
Protein crystallographic studies to understand the reaction mechanism of enzymes: α-methylacyl-CoA racemase and argininosuccinate lyase
P. Bhaumik (2006)
A mouse model for alpha-methylacyl-CoA racemase deficiency: adjustment of bile acid synthesis and intolerance to dietary methyl-branched lipids.
K. Savolainen (2004)
Effect of clofibrate on the chiral disposition of ibuprofen in rats.
S. Scheuerer (1998)
A novel colorimetric assay for α-methylacyl-CoA racemase 1A (AMACR; P504S) utilizing the elimination of 2,4-dinitrophenolate.
Maksims Yevglevskis (2017)
Stereoisomeric bio‐inversion key to biosynthesis of firefly d‐luciferin
K. Niwa (2006)
α-Methylacyl-CoA racemase : an enzyme at crossroads in lipid metabolism
K. Savolainen (2004)
Can p503s, p504s and p510s gene expression in peripheral-blood be useful as a marker of prostatic cancer?
M. Cardillo (2004)
α‐Methylacyl‐CoA racemase – an ‘obscure’ metabolic enzyme takes centre stage
M. Lloyd (2008)
Role of racemization in optically active drugs development.
I. Ali (2007)
The stereo inversion of 2‐arylpropionic acid non‐steroidal anti‐inflammatory drugs and structurally related compounds by Verticillium lecanii
Thomason (1998)
Cellular signaling, AGE accumulation and gene expression in hepatocytes of lean aging rats fed ad libitum or food-restricted
L. Teillet (2002)
Metabolic Chiral Inversion of 2-Arylpropionic Acids
I. Tegeder (2003)
Enantioselective Pharmacokinetics of Ibuprofen and Involved Mechanisms
H. Hao (2005)
Syntheses and cytotoxicity of phosphatidylcholines containing ibuprofen or naproxen moieties
Marek Kłobucki (2019)
Identification of differentially expressed genes in human prostate cancer using subtraction and microarray.
J. Xu (2000)
Ibuprofen: Pharmacology, Therapeutics and Side Effects
K. Rainsford (2012)
Biochemistry and genetics of inherited disorders of peroxisomal fatty acid metabolism
P. Veldhoven (2010)
Chiral separation of ibuprofen and chiral pharmacokinetics in healthy chinese volunteers
Chaonan Zheng (2010)
Crystal structure of fatty acid‐CoA racemase from Mycobacterium tuberculosis H37Rv
K. S. Lee (2006)
Microbial Deracemization of α-Substituted Carboxylic Acids
D. Kato (2002)
Influence of CYP2C8 and CYP2C9 polymorphisms on pharmacokinetic and pharmacodynamic parameters of racemic and enantiomeric forms of ibuprofen in healthy volunteers.
R. López-Rodríguez (2008)
Chemistry around imidazopyrazine and ibuprofen: discovery of novel fatty acid amide hydrolase (FAAH) inhibitors.
Frédéric De Wael (2010)
Clinical Pharmacokinetics of Ibuprofen
N. M. Davies (1998)
Evidence of COX-2 independent induction of apoptosis and cell cycle block in human colon carcinoma cells after S- or R-ibuprofen treatment.
A. Janssen (2006)
Variants in the α-Methylacyl-CoA Racemase Gene and the Association with Advanced Distal Colorectal Adenoma
Sarah E. Daugherty (2007)
α-Methylacyl-CoA racemase (AMACR): metabolic enzyme, drug metabolizer and cancer marker P504S.
M. Lloyd (2013)
Synthesis and use of isotope-labelled substrates for a mechanistic study on human alpha-methylacyl-CoA racemase 1A (AMACR; P504S).
D. Darley (2009)
The bile acid-inducible baiF gene from Eubacterium sp. strain VPI 12708 encodes a bile acid-coenzyme A hydrolase.
H. Q. Ye (1999)
A combined experimental and computational study of the molecular interactions between anionic ibuprofen and water.
Andy D. Zapata-Escobar (2014)
See more
Semantic Scholar Logo Some data provided by SemanticScholar