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. Author manuscript; available in PMC: 2011 Mar 24.
Published in final edited form as: J Am Chem Soc. 2010 Mar 24;132(11):3914–3922. doi: 10.1021/ja910578y

Figure 5.

Figure 5

COX inhibitors attenuate ONOO--induced nitration and prevent conformational changes in COX-1. In all reactions, COX-1 complexes were at 0.21 mg/ml and ONOO- was at 250 μM (a) Quantification of total 3-NT by HPLC-EC for the reaction of FePPCOX-1 with ONOO- ± arachidonic acid (AA; 100 μM) and the COX inhibitors aspirin (70 μM) and indomethacin (14 μM). Data are averages ± SEM. *p ≤ 0.029 for AA-bound FePPCOX-1 relative to substrate-free FePPCOX-1. #p ≤ 0.002 for aspirin-treated FePPCOX-1 relative to AA-bound FePPCOX-1. †p ≤ 0.001 for indomethacin-treated FePPCOX-1 relative to AA-bound FePPCOX-1. (b) Western blots for the reaction of FePPCOX-1 with ONOO- ± AA (100 μM) and the COX inhibitors aspirin (70 μM) and indomethacin (14 μM). Blots were probed with monoclonal anti-3-NT (top) and with polyclonal anti-3-nitro-Tyr385-peptide in COX-1 (bottom). (c) CD spectra recorded in the far UV region at 25°C compare the structure of FePPCOX-1 (black) to aspirin-treated FePPCOX-1 following reaction with ONOO-.