Skip to main content
. 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 3.

Figure 3

Arachidonic acid (AA) binding redirects ONOO--induced COX-1 nitration. In all reactions, COX-1 complexes were at 0.21 mg/ml, AA was at 100 μM and ONOO- was at 250 μM. (a) Quantification of total 3-NT by HPLC-EC for the reaction of FePPCOX-1 and CoPPCOX-1 with ONOO-. Total 3-NT for the reaction of AA-bound CoPPCOX-1 with ONOO- was also quantified. Data are averages ± SEM. *p ≤ 0.023 for FePPCOX-1 relative to CoPPCOX-1. #p ≤ 0.035 for AA-bound CoPPCOX-1 relative to FePPCOX-1. (b) Western blots compare the reactions of FePPCOX-1 and CoPPCOX-1 with ONOO- ± AA. Blots were probed with monoclonal anti-3-NT and with (c) polyclonal anti-3-nitro-Tyr385-peptide in COX-1. (d) Quantification of total 3-NT by HPLC-EC for the reaction of FePPCOX-1 with ONOO- ± AA and eicosapentanoic acid (EPA; 100 μM). Data are averages ± SEM. *p ≤ 0.027 for AA-bound FePPCOX-1 relative to substrate-free FePPCOX-1. #p ≤ 0.026 for EPA-bound FePPCOX-1 relative to AA-bound FePPCOX-1. (e) Western blots for the reaction of FePPCOX-1 with ONOO- ± AA and EPA. Blots were probed similarly to (b) and (c).