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. Author manuscript; available in PMC: 2016 Aug 3.

Published in final edited form as: FEBS J. 2015 Aug 3;282(16):3199–3217. doi: 10.1111/febs.13359

Fig. 3 — Determination of the pK_a value (Panel A) and changes of the absorption spectra (Panels B-E) for the difluoromethyl derivative 9 (BJ735) upon glutathione addition. (B) Absorption spectrophotometric variation and (C) Absorbance at 350 nm measured in the time course of the glutathionylation reaction with the difluoromethyl derivative 9. Solvent: 200μL MeCN + 265 μL NH₄OH buffer at pH 7.5; [9] = 2 × 10⁻⁴ M (10 μL stock at 10 mM); [GSH] = 10⁻³ M (25 μL stock at 20 mM); T = 25°C. (1) t = 0 s; (2) t = 1800 s. (D) Electronic absorption spectra measured for the difluoromethyl derivative 9 and its glutathione adducts and (E) Distribution diagram of the difluoromethyl derivative 9 and its products as a function of time upon the glutathionylation reaction. Solvent: 200μL MeCN + 265 μL NH₄OH buffer at pH 7.5; [9] = 2 × 10⁻⁴ M (10 μL stock at 10 mM); [GSH] = 10⁻³ M (25 μL stock at 20 mM); T = 25°C.

Fig. 3 — Determination of the pK_a value (Panel A) and changes of the absorption spectra (Panels B-E) for the difluoromethyl derivative 9 (BJ735) upon glutathione addition. (B) Absorption spectrophotometric variation and (C) Absorbance at 350 nm measured in the time course of the glutathionylation reaction with the difluoromethyl derivative 9. Solvent: 200μL MeCN + 265 μL NH₄OH buffer at pH 7.5; [9] = 2 × 10⁻⁴ M (10 μL stock at 10 mM); [GSH] = 10⁻³ M (25 μL stock at 20 mM); T = 25°C. (1) t = 0 s; (2) t = 1800 s. (D) Electronic absorption spectra measured for the difluoromethyl derivative 9 and its glutathione adducts and (E) Distribution diagram of the difluoromethyl derivative 9 and its products as a function of time upon the glutathionylation reaction. Solvent: 200μL MeCN + 265 μL NH₄OH buffer at pH 7.5; [9] = 2 × 10⁻⁴ M (10 μL stock at 10 mM); [GSH] = 10⁻³ M (25 μL stock at 20 mM); T = 25°C.