S-Glutathionylation of serum and liver proteins in GSTπ WT
and KO mice. Male GSTπ wild-type (GSTP1P2+/+) and knock-out
(GSTP1P2-/-) mice (ages 1 and 3 months) were treated with an
intravenous bolus of the oxidized glutathione mimetic, NOV-002 at 25 mg/kg or
PABA/NO at 5 mg/kg. Blood was collected at various time points via orbital
bleed. The plasma proteins (A) were separated by non-reducing
SDS-PAGE and S-glutathionylated contrapsin was evaluated by
immunoblot (IB) with PSSG antibody. The ratios of
S-glutathionylated contrapsin to albumin were plotted in arbitrary
units (B), solid, 1-month-old; hatched, 3-month-old
animals (6 animals per treatment group). Free protein sulfhydryl content was
measured in the liver homogenates of 3-month-old GSTπ wild-type (+/+) and
knock-out (-/-) mice (C) using the fluorescent sulfhydryl-specific
probe ThioGlo-1. The ThioGlo-1 emission (at 513 nm) for each treatment group
was averaged and plotted as mean ± S.D. (n = 3); solid
bars represent GSTπ WT (+/+) and open bars represent KO (-/-)
animals. The difference between WT and KO treated and untreated animals is
statistically significant (*, p ≤ 0.001).