Techniques used to characterize the binding of cyclooxygenase inhibitors to the cyclooxygenase active site.
Inhibitors of enzyme-catalyzed reactions are typically characterized by their ability to diminish product formation while altering the Michaelis Menten constants V(max) and K(m). Determination of an apparent inhibitor affinity (K(i)) for the enzyme is also possible using this approach. Unfortunately, analysis of product formation does not easily provide information regarding the kinetics of inhibitor binding and may not be possible depending upon the mechanism of action. Radiolabeling of the inhibitor allows one to do a direct binding assay and thereby more directly determine the kinetics of inhibitor binding. With this in mind, we developed a radioligand-based binding assay for inhibitors of cyclooxygenase.
Hood
William F
WF
Pfizer Global Research and Development, Chesterfield, MO, USA.
D020134:Catalytic Domain; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology*; D004800:Enzymes, Immobilized / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism*; D011485:Protein Binding; D011869:Radioligand Assay / Q000379:methods*
D016861:Cyclooxygenase Inhibitors; D004800:Enzymes, Immobilized; D011451:Prostaglandin-Endoperoxide Synthases