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. 2019 Oct 31;20(21):5440. doi: 10.3390/ijms20215440

Figure 1.

Figure 1

Reduction of the RNase and reaction of the rRNase with DTNB. The reduction of the RNase (0.05 mM) was performed in 10 mM borate buffer, pH 8.5, with 0.5 mM DTT and urea 8 M at two different temperatures 37 °C (A) and 60 °C (B) (fit obtained using Equation (1)). The reduced cysteines were titrated at different times as reported under Methods. Average ± SD derived from ten independent experiments. (C) 1.25 nmol of the rRNase (about 10 nmol titratable cysteines) was reacted with 20 nmol DTNB in 1 mL of 0.1 M sodium acetate buffer, pH 5.0 and 0.2 M urea (red line). For comparison, an identical reaction was performed with 10 nmol of GSH (black line). (D) First-order kinetic constants for the reaction of the eight cysteines of the rRNase with different DTNB concentrations at pH 5.0 and 0.2 M urea. Average ± SD derived from three independent experiments.