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. 2024 Dec 4;68(4):467–478. doi: 10.1042/EBC20230095

Figure 2. Schematic overview of postulated modified biotin-switch assay.

Figure 2

Schematic overview of postulated modified biotin-switch assay proposed by Mustafa et al. [35]. MMTS was postulated to solely labelled free thiols, followed by the alkylation of –SSH group by biotin-HPDP. Streptavidin beads were employed to covalently pull down biotinylated-persulfidated proteins. The use of DTT cleaved the disulfide bond (-R-S-S-Biotin), thus eluting the persulfidated proteins. However, in addition to the interference of intra- or intermolecular disulfides, MMTS reacts faster with persulfides than free thiols, a key caveat of this method. Biotin-HPDP, N-[6-(biotinamido)hexyl]-3′-(2-pyridyldithio) propionamide; DTT, dithiothreitol; MMTS, S-methyl methanethiosulfonate (Created with BioRender.com.)

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