Fig. 7: Acetylation and deacetylation of Ub / Ub AcK variants by p300 and HDAC6, respectively.

a Monomeric Ub is acetylated at K48 by p300 in presence of acetyl-CoA (Ub), while only a background signal can be observed in the absence of either acetyl-CoA (Ub w/o AcCoA) or p300 (Ub w/o p300). b M1-linked Ub dimers are acetylated at K11 and K48 by p300 in presence of acetyl-CoA (diUb), while in its absence (diUb w/o AcCoA) or in the absence of p300 (diUb w/o p300) only background signals are observed. a, b Ub acetylation was normalized against a highly abundant C-terminal peptide of Ub (aa 64–73). ++ and +++ indicate doubly and triply protonated charge states of the detected peptide, respectively. For quantification, the respective product ions (MS2-level) of the indicated precursors were used. Error bars indicate SEM of three independent measurements. c HDAC6 deacetylates the various Ub AcK variants with similar efficiency. Ub AcK variants were incubated with HDAC6 and reaction products were analyzed by intact protein MS. Peak areas of the respective acetylated and deacetylated Ub variants were used for quantification of deacetylation and normalized to the total Ub peak area. Data were obtained in two independent experiments. Source data are provided as a Source Data file.