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. 2017 Nov 27;2(11):8313–8318. doi: 10.1021/acsomega.7b01318

Figure 1.

Figure 1

Inhibition and reactivation of the steady-state activity of SHP2 variants. (A) Domain cartoon of the SHP2cat construct. (B) Domain cartoon of the SHP21–526 construct (SH2N, cyan; SH2C, blue; CAT, orange). (C, D) The steady-state activity of SHP2 variants (C, SHP2237-529; D, SHP21–526; WT, black squares, N308D variant, red diamonds) were measured after incubating the samples with the indicated concentrations of H2O2 for 15 min. The % remaining activity was calculated by normalizing the measured activity to the sample incubated without H2O2. (E, F) The ability of oxidized SHP2 variants (E, SHP2237-529; F, SHP21–526) to be subsequently reactivated was determined by measuring the activities of reduced (no H2O2) and oxidized (15 min incubation with 500 μM H2O2 prior to activity measurements) SHP2 samples in the presence of the indicated concentrations of DTT. The % reactivation was calculated by normalizing the activities of the oxidized samples to the corresponding reduced samples. ±SE, n = 3–4. Two-way ANOVA test, with ****p < 0.0001 (C, D) or *p < 0.05 (F) between WT and mutant.