In Vitro Biochemical Analysis of Recombinant Plant Proteins Under Oxidation

AbstractBiochemical analysis is crucial for determining protein functionality changes during various conditions, including oxidative stress conditions. In this chapter, after giving brief guidelines for experimental design, we provide step-by-step instructions to purify recombinant plant proteins from E. coli, to prepare reduced and oxidized proteins for activity assay, and to characterize the protein under reducing and oxidizing conditions, with a focus on thiol-based oxidative modifications, like S-sulfenylation and disulfide formations.Key wordsRecombinant protein expressionProtein purificationOxidation and reductionPost-translational modificationsS-sulfenylation detection