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Dop has a broad substrate specificity with respect to pupylated proteins. Pupylated proteins (300 μg) isolated from Msm Δdop expressing amino-terminally His6-tagged Pup-GGE were incubated with ATP (5 mM) in the presence or absence of Dop (1 μM). Samples were withdrawn at the time points indicated and analysed by an anti-Pup immunoblot. Dop, deamidase of Pup; Pup, prokaryotic ubiquitin-like protein.
