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. 2019 Oct 24;294(49):18863–18872. doi: 10.1074/jbc.RA119.011237

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© 2019 Andreeva et al.

Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 7. — GSH-MGO hemithioacetal is not a substrate for DJ-1. GSH and MGO at the indicated concentrations were mixed in PBS to form GSH-MGO and incubated with 20 μm DJ-1 at 30 °C. A, kinetic curves of GSH-MGO decomposition by DJ-1 were obtained using optical absorbance at 288 nm as described in supporting methods. B, concentrations of free MGO were calculated for the data sets shown in A, assuming a fast equilibrium between MGO and GSH-MGO. In the case where MGO and GSH-MGO are initially present in comparable amounts (black curves), the decrease in GSH-MGO is accompanied by a significant decrease in free MGO due to fast equilibrium. C is a sum of the curves in A and B, which corresponds to the kinetic profile of total MGO (free + hemithioacetal) in both samples. Note that the concentration of total MGO decreases faster in the case of a lower initial concentration of GSH-MGO (black curves), a result inconsistent with GSH-MGO being a substrate of DJ-1.