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. 2016 May 20;291(29):15069–15081. doi: 10.1074/jbc.M116.725713

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© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 7. — Fenamic acid inhibitors quench preformed huCOX-2 tyrosyl radicals. Tyrosyl radicals were generated by reacting reconstituted huCOX-2 with hydrogen peroxide in ice for 30 s. Varying concentrations of mefenamic acid (A) and tolfenamic acid (B) were then added and mixed for 2–3 s before freezing the sample and collecting ESR spectra. The arrows indicate the decrease in ESR spectra amplitude as the inhibitor concentration is increased. C, calculation of the total radical yield, based on double integration of the spectra in A and B. The double integral of the spectrum with no added inhibitor was normalized to 100% radical yield.