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Figure 4
Oligomerization states of VvPrx3 in solution were analyzed using size-exclusion chromatography, and fractions were further analyzed by SDS–PAGE under nonreducing conditions. The VvPrx3 (C73S) protein (33 µM) was incubated with H2O2 (15 µM) for 60 min before applying size-exclusion chromatography. The result of the native gel electrophoresis is provided in Supplementary Fig. S5. The molecular size was calculated from the standard plot in Supplementary Fig. S8. The molecular sizes of the fractions labelled (a)–(d) correspond to decamer (∼244 kDa), hexamer (∼143 kDa), tetramer (∼81 kDa) and dimer (∼42 kDa), respectively. The protein band of disulfide-bond-containing VvPrx3 (D) and bands that lack a disulfide bond (M) are indicated. As a control for the non-oxidized sample, VvPrx3 (C73S/C48S) was analyzed using the same column (shown by a broken line).

IUCrJ
Volume 5| Part 1| January 2018| Pages 82-92
ISSN: 2052-2525
https://doi.org/10.1107/S205225251701750X