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. Author manuscript; available in PMC: 2012 Feb 8.
Published in final edited form as: Biochemistry. 2011 Jan 12;50(5):727–737. doi: 10.1021/bi101355a

Figure 3. picNuA4 randomly acetylates free and nucleosomal H4.

Figure 3

A) picNuA4 acetylation reactions, in which limiting amounts of AcCoA were added to reactions and visualized by SYPRO Ruby stained TAU electrophoresis. Acetylation bands were excised, deuterated with acetic anhydride and analyzed by LC-MS/MS. Graphs depict percent picNuA4-acetylated peptide in free H4 (B), and H4 within the NCPs (C), plotted by lysine position and acetylation state. Reaction conditions: 1 μM picNuA4, 10 μM H4 or 2 μM NCP in 50 mM Tris (pH 7.5) and 1 mM DTT. Data sets are representative of n = 3 for H4 and n = 1 for NCP experiments. MS experimental replicates differed no more than 6% per acetylation site for H4 (see Supplemental Figure 2D).