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. Author manuscript; available in PMC: 2023 May 5.
Published in final edited form as: Structure. 2022 Mar 25;30(5):763–776.e4. doi: 10.1016/j.str.2022.03.002

Figure 5, related to Figure S5 and S6. Cysteine oxidation and dimerization/aggregation of aged samples.

Figure 5,

(A) Assessment of the decrease in free thiol content over time using Ellman’s assay for γS-wt (black), γS-3 (yellow), γS-5 (blue), γS-7 (green), and γS-9 (pink). Measurements were performed in triplicate. Data are represented as mean ± SD. (B) Size-exclusion chromatography in line with multiangle light scattering (SEC-MALS) conducted on each variant after aging for 1 month. High molecular weight (HMW) peaks are seen in γS-5, γS-7, γS-9. (C) Intact mass spectra of each aged variant after reaction with iodoacetamide (IAM) and subsequent reduction with dithiothreitol (DTT). The average free thiol content per protein molecule weighted by the peak intensity decreases with increasing deamidation.