Grx2−/− alters 2GSH/GSSG redox potential and the glutathionylated proteome but does not induce oxidative damage.
a, measurement of the absolute levels of GSH and GSSG in liver and muscle mitochondria from WT and Grx2−/− mice. 2GSH/GSSG was calculated from the absolute GSH and GSSG levels. Eh was calculated using the Nernst equation as described under “Materials and Methods.” ** indicates p ≤ 0.01. Student's t test, n = 4, mean ± S.E. b, serum levels of GSH and GSSG in WT and Grx2−/− mice. n = 4, mean ± S.E., Student's t test. c, measurement of HNE-His adducts in liver and muscle mitochondria from WT and Grx2−/− mice. n = 4, mean ± S.E., Student's t test. d, assessment of the glutathionylated proteome in liver and muscle mitochondria. Gels were run under nonreducing conditions to preserve PSSG adducts. Samples prepared in 2% (v/v) β-mercaptoethanol served as the control. Blots were quantified using ImageJ software. Student's t test, n = 5, mean ± S.E. e, immunodetection of glutaredoxin-2, Grx2; thioredoxin-2, Trx2; and uncoupling protein-3, UCP3. SDH, succinate dehydrogenase.