skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Competition and allostery govern substrate selectivity of cyclooxygenase-2

Journal Article · · Proceedings of the National Academy of Sciences of the United States of America
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1]
  1. Vanderbilt Univ., Nashville, TN (United States)
+ Show Author Affiliations

Cyclooxygenase-2 (COX-2) oxygenates arachidonic acid (AA) and its ester analog, 2-arachidonoylglycerol (2-AG), to prostaglandins (PGs) and prostaglandin glyceryl esters (PG-Gs), respectively. While the efficiency of oxygenation of these substrates by COX-2 in vitro is similar, cellular biosynthesis of PGs far exceeds that of PG-Gs. Evidence that the COX enzymes are functional heterodimers suggests that competitive interaction of AA and 2-AG at the allosteric site of COX-2 might result in differential regulation of the oxygenation of the two substrates when both are present. Modulation of AA levels in RAW264.7 macrophages uncovered an inverse correlation between cellular AA levels and PG-G biosynthesis. In vitro kinetic analysis using purified protein demonstrated that the inhibition of 2-AG oxygenation by high concentrations of AA far exceeded the inhibition of AA oxygenation by high concentrations of 2-AG. An unbiased systems-based mechanistic model of the kinetic data revealed that binding of AA or 2-AG at the allosteric site of COX-2 results in a decreased catalytic efficiency of the enzyme toward 2-AG, whereas 2-AG binding at the allosteric site increases COX-2’s efficiency toward AA. The results suggest that substrates interact with COX-2 via multiple potential complexes involving binding to both the catalytic and allosteric sites. Competition among AA and 2-AG for these sites, combined with differential allosteric modulation, gives rise to a complex interplay between the substrates, leading to preferential oxygenation of AA.

Research Organization:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States). Oak Ridge Leadership Computing Facility (OLCF)
Sponsoring Organization:
USDOE Office of Science (SC); National Institutes of Health (NIH); National Science Foundation (NSF)
OSTI ID:
1565482
Journal Information:
Proceedings of the National Academy of Sciences of the United States of America, Vol. 112, Issue 40; ISSN 0027-8424
Publisher:
National Academy of SciencesCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 22 works
Citation information provided by
Web of Science

References (35)

Programming biological models in Python using PySB journal January 2013
(R)-Profens are substrate-selective inhibitors of endocannabinoid oxygenation by COX-2 journal September 2011
13-Methylarachidonic Acid Is a Positive Allosteric Modulator of Endocannabinoid Oxygenation by Cyclooxygenase journal February 2015
Regulation of arachidonic acid release and cytosolic Phospholipase A 2 activation journal March 1999
The Structural Basis of Endocannabinoid Oxygenation by Cyclooxygenase-2 journal April 2011
Global Kinetic Explorer: A new computer program for dynamic simulation and fitting of kinetic data journal April 2009
Cytosolic Phospholipase A 2 Is Required for Macrophage Arachidonic Acid Release by Agonists That Do and Do Not Mobilize Calcium : NOVEL ROLE OF MITOGEN-ACTIVATED PROTEIN KINASE PATHWAYS IN CYTOSOLIC PHOSPHOLIPASE A journal April 2000
Lipid Profiling Reveals Arachidonate Deficiency in RAW264.7 Cells:  Structural and Functional Implications † journal December 2006
PyMC : Bayesian Stochastic Modelling in Python journal January 2010
FitSpace Explorer: An algorithm to evaluate multidimensional parameter space in fitting kinetic data journal April 2009
Human Cyclooxygenase-2 Is a Sequence Homodimer That Functions as a Conformational Heterodimer journal April 2011
Localization and regulation of cytosolic phospholipase A2 journal October 2000
Prostaglandin H Synthase-2-catalyzed Oxygenation of 2-Arachidonoylglycerol Is More Sensitive to Peroxide Tone than Oxygenation of Arachidonic Acid journal September 2012
Mechanism of Free Radical Oxygenation of Polyunsaturated Fatty Acids by Cyclooxygenases journal June 2003
Structural Basis of Fatty Acid Substrate Binding to Cyclooxygenase-2 journal May 2010
Differential Evolution Markov Chain with snooker updater and fewer chains journal October 2008
Prostaglandins and Leukotrienes: Advances in Eicosanoid Biology journal November 2001
Phospholipase A 2 inhibitors as potential therapeutic agents for the treatment of inflammatory diseases journal December 2009
Endocannabinoid Oxygenation by Cyclooxygenases, Lipoxygenases, and Cytochromes P450: Cross-Talk between the Eicosanoid and Endocannabinoid Signaling Pathways journal October 2011
Human cyclooxygenase-1 activity and its responses to COX inhibitors are allosterically regulated by nonsubstrate fatty acids journal April 2012
Properties of cell death models calibrated and compared using Bayesian approaches journal January 2013
Metabolism of the Endocannabinoids, 2-Arachidonylglycerol and Anandamide, into Prostaglandin, Thromboxane, and Prostacyclin Glycerol Esters and Ethanolamides journal September 2002
Glycerylprostaglandin Synthesis by Resident Peritoneal Macrophages in Response to a Zymosan Stimulus journal May 2005
Oxygenation of the Endocannabinoid, 2-Arachidonylglycerol, to Glyceryl Prostaglandins by Cyclooxygenase-2 journal August 2000
Hydroperoxide Dependence and Cooperative Cyclooxygenase Kinetics in Prostaglandin H Synthase-1 and -2 journal July 1999
Analysis of endocannabinoids by Ag+ coordination tandem mass spectrometry journal March 2003
Simultaneous analysis of prostaglandin glyceryl esters and prostaglandins by electrospray tandem mass spectrometry journal August 2005
Synthesis of Prostaglandin E 2 Ethanolamide from Anandamide by Cyclooxygenase-2 journal August 1997
Indole Cytosolic Phospholipase A 2 α Inhibitors: Discovery and in Vitro and in Vivo Characterization of 4-{3-[5-Chloro-2-(2-{[(3,4-dichlorobenzyl)sulfonyl]amino}ethyl)-1-(diphenylmethyl)-1 H -indol-3-yl]propyl}benzoic Acid, Efipladib journal June 2008
The Binding of Arachidonic Acid in the Cyclooxygenase Active Site of Mouse Prostaglandin Endoperoxide Synthase-2 (COX-2): A PUTATIVE L-SHAPED BINDING CONFORMATION UTILIZING THE TOP CHANNEL REGION journal August 1999
Zymosan-induced glycerylprostaglandin and prostaglandin synthesis in resident peritoneal macrophages: roles of cyclo-oxygenase-1 and -2 journal September 2006
Differential Allosteric Regulation of Prostaglandin H Synthase 1 and 2 by Arachidonic Acid journal May 1997
Differential Sensitivity and Mechanism of Inhibition of COX-2 Oxygenation of Arachidonic Acid and 2-Arachidonoylglycerol by Ibuprofen and Mefenamic Acid journal August 2009
Cytosolic phospholipase A2 is essential for both the immediate and the delayed phases of eicosanoid generation in mouse bone marrow-derived mast cells journal April 1999
Evaluating the Accuracy of Sampling-Based Approaches to the Calculation of Posterior Moments posted_content January 1991

Cited By (7)

Signal integration and information transfer in an allosterically regulated network journal July 2019
Dual cyclooxygenase–fatty acid amide hydrolase inhibitor exploits novel binding interactions in the cyclooxygenase active site journal January 2018
PyDREAM: high-dimensional parameter inference for biological models in python journal October 2017
Inhibition of key enzymes in the inflammatory pathway by hybrid molecules of terpenes and synthetic drugs: In vitro and in silico studies journal October 2018
A New Look at the Structures of Old Sepsis Actors by Exploratory Data Analysis Tools journal November 2019
Signal integration and information transfer in an allosterically regulated network posted_content January 2019
Conservative Secondary Shell Substitution In Cyclooxygenase-2 Reduces Inhibition by Indomethacin Amides and Esters via Altered Enzyme Dynamics journal December 2015

Similar Records

Dual cyclooxygenase–fatty acid amide hydrolase inhibitor exploits novel binding interactions in the cyclooxygenase active site
Journal Article · Thu Mar 01 00:00:00 EST 2018 · Journal of Biological Chemistry · OSTI ID:1565482
+5 more
Comparison of Cyclooxygenase-1 Crystal Structures: Cross-Talk between Monomers Comprising Cyclooxygenase-1 Homodimers
Journal Article · Mon Nov 01 00:00:00 EDT 2010 · Biochemistry-US · OSTI ID:1565482
+1 more
Coxibs interfere with the action of aspirin by binding tightly to one monomer of cyclooxygenase-1
Journal Article · Thu Feb 11 00:00:00 EST 2010 · Proc. Natl. Acad. Sci. USA · OSTI ID:1565482
+7 more

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
cyclooxygenase
endocannabinoids
allosteric regulation
chemical kinetics
Bayesian inference