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. 2009 Nov 23;285(5):3168–3180. doi: 10.1074/jbc.M109.056846

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 4. — Immunoinhibition of NQR-mediated generation activity by AbGSCA206 and AbGSCB367 as assayed by EPR spin-trapping with DEPMPO. A, titration of NQR-mediated generation by antibodies of AbGSCA206 (○), AbGSCB367 (△), and pre-immune IgG (▼). Isolated NQR (0.91 mg/ml) was incubated with various amounts of antibodies at 4 °C for 12 h. The generation mediated by NQR was initiated by NADH (0.5 mm) in the presence of DEPMPO (20 mm), and subsequently measured by EPR as described under “Experimental Procedures.” B–D, the computer simulation (dashed line) superimposed on the experimental EPR spectra (solid line) obtained from the reaction systems containing NQR (0.91 mg/ml), antibodies (0.36 mg/ml, pre-immune IgG in B, AbGSCA206 in C, and AbGSCB367 in D), diethylenetriaminepentaacetic acid (1 mm), DEPMPO (20 mm), and NADH (0.5 mm). B′, C′, and D′, same as B, C, and D except that Mn-SOD (1 unit/μl) was present in the reaction mixture.

Inline graphic — Immunoinhibition of NQR-mediated generation activity by AbGSCA206 and AbGSCB367 as assayed by EPR spin-trapping with DEPMPO. A, titration of NQR-mediated generation by antibodies of AbGSCA206 (○), AbGSCB367 (△), and pre-immune IgG (▼). Isolated NQR (0.91 mg/ml) was incubated with various amounts of antibodies at 4 °C for 12 h. The generation mediated by NQR was initiated by NADH (0.5 mm) in the presence of DEPMPO (20 mm), and subsequently measured by EPR as described under “Experimental Procedures.” B–D, the computer simulation (dashed line) superimposed on the experimental EPR spectra (solid line) obtained from the reaction systems containing NQR (0.91 mg/ml), antibodies (0.36 mg/ml, pre-immune IgG in B, AbGSCA206 in C, and AbGSCB367 in D), diethylenetriaminepentaacetic acid (1 mm), DEPMPO (20 mm), and NADH (0.5 mm). B′, C′, and D′, same as B, C, and D except that Mn-SOD (1 unit/μl) was present in the reaction mixture.