Figure 6.

Succinylated Abf2p is less sensitive to digestion by the ATP-dependent ScLon protease. Nonsuccinylated (Abf2p) or in vitro–succinylated (succ-Abf2p) protein was incubated with purified ScLon in the presence or absence of a dsDNA probe. A, the course of an Abf2p ScLon-mediated digestion in reaction mixtures containing 2 mM ATP and 10 mM MgCl₂ on Coomassie Brilliant Blue–stained gels. The reactions were terminated at the indicated times and the samples loaded onto a 12% SDS-polyacrylamide gel. B, quantification of the digestion experiments shown in panel A. Band intensity evaluation of undigested Abf2p (Abf2 (%)) was performed with ImageJ (v.1.53) (104) separately for each studied protein. The height of a given bar represents the mean value, and the error bars show ± one SD from at least three separate measurements. C, the course of β-casein digestion by ScLon. β-casein was incubated with ScLon in the presence or absence of a dsDNA in a reaction mixture containing 2 mM ATP and 10 mM MgCl₂ for indicated times, and the proteins were separated on a 12% SDS-polyacrylamide gel.