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. 2008 Jun 27;283(26):17898–17907. doi: 10.1074/jbc.M710609200

FIGURE 5.

FIGURE 5.

Hysteretic behavior of GR activity of TGR. Full-time courses obtained using different assay conditions are shown. In all cases the reactions were started by the addition of TGRGCUG at the indicated final concentrations. A, effect of enzyme concentration. Assays were performed at varying TGRGCUG concentrations and constant NADPH and GSSG concentrations (100 μm and 1 mm, respectively). B, effect of GSSG concentration. GSSG concentration was varied at constant NADPH and enzyme concentrations (100 μm and 10 nm, respectively). Note that at 31 and 62 μm GSSG reactions come to their end at higher A340, i.e. before depleting NADPH, because GSSG becomes the limiting reagent. C, effect of GSH concentration. GSH was included at various concentrations while maintaining constant GSSG, NADPH, and enzyme concentrations (1 mm, 125 μm, and 10 nm, respectively). D, effect of Trx concentration. cTrx was added at different concentrations to reaction mixtures containing 1 mm GSSG, 125 μm NADPH, and 10 nm TGRGCUG. The TGRGCUG concentration considered was corrected according to its selenium content.