Figure 6. The synthesis of H3K4su and its application to test the desuccinylation activity of Sirt5.
(A) Using dPPMT-NB-Su to convert H3K4AznL to H3K4su. Reaction conditions and analysis were as same as for the synthesis of Ub-K48su. (B) The tandem MS analysis of the trypsinized Ksu-containing fragment of the 48 h reaction and UV treated product of H3K4AznL. (C) Desuccinylation of the H3K4su-H4 tetramer by Sirt1 and Sirt5. 180 μM H3K4su-H3 tetramer was incubated with or without 0.5 μM Sirt1/Sirt5 in the presence of 10 mM NAD+ for 4 h. After reactions, proteins were analyzed in the Western blot analysis using anti-H3 and pan anti-Ksu antibodies.