Fig. 7.

Deacylation analysis using ScCobB1 and ScCobB2 in vitro. The regulatory crosstalk between deacetylation and desuccinylation was tested by treating the same synthetic peptides with sirtuins. The assays were performed as explained in the Experimental procedures section, using NAD⁺ as the cofactor. The assays without NAD⁺ in the mixtures were used as the negative controls (see supplemental Fig. S5 for more details). A, the modified peptides were incubated with ScCobB1. Relatively, low deacetylase and desuccinylase activities were observed. B, the modified peptides were incubated with ScCobB2. A specific desuccinylase activity was observed. C, the bifunctional CobBL was used as a positive control that showed both the deacetylase and desuccinylase activities.