Fig. 8.

Alb and GSH/GSSG interaction reciprocally regulate the activity of -SH groups. (A) Effect of GSH and GSSG on the number of –SH groups in the differently modified Alb. Differently modified Alb was allowed to react with 2 mM GSH or GSSG overnight at 4 °C. After removing the unreacted GSH and GSSG, the equal number of treated samples was assayed for SH groups using the fluorescent maleimide-labeling assay. The samples were separated with 10% SDS-PAGE and detected for the fluorescent signal. The equal loading of proteins was confirmed by staining the gel with EZ blue. Note the changes in the fluorescent intensity of bands and the shift of bands after treatment with GSH or GSSG. (B) Influence of the modified Alb on GSH and GSSG ratio. A mixture of GSH and GSSG at the estimated concention of 5 μM was incubated with different forms of Alb (0.1 mg/ml in 100 μl distilled water) for 60 min and assayed for GSH and GSSG concentrations. Data shown are mean ± SD (n = 4; ^##P < 0.01 vs. DW; *P < 0.05, **P < 0.01). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)