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. 2011 Jun 10;286(33):29098–29107. doi: 10.1074/jbc.M111.240127

FIGURE 5.

FIGURE 5.

Glutathione and DMPO compete for O2˙̄-induced eNOS protein radical. a, GSH dose-dependently inhibits O2˙̄-induced eNOS DMPO adduct. The upper panel is a blot against anti-DMPO antibody. The lower panel is a blot against anti-eNOS antibody. Lane 1, BH4-free eNOS in the presence of all NOS cofactors and 50 mm DMPO, and the reaction is initiated with 1 mm NADPH for 30 min at room temperature. Lane 2, the reaction is the same as in lane 1 with the addition of 1 mm GSH. Lane 3, with addition of 5 mm GSH. Lane 4, with addition of 10 mm GSH. b, DMPO dose-dependently inhibits O2˙̄-induced eNOS S-glutathionylation. The upper panel is a blot against anti-GSH antibody. The lower panel is a blot against anti-eNOS antibody. Lane 1, BH4-free eNOS in the presence of all NOS cofactors and 2 mm GSH, with the reaction initiated by 1 mm NADPH for 30 min at room temperature. Lane 2, the reaction is the same as in lane 1 with the addition of 20 mm DMPO. Lane 3, with the addition of 50 mm DMPO. Lane 4, with the addition of 100 mm DMPO.