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Synthesis of acetylated H4 peptides to define the substrate specificity of an HDAC. (a) Limited proteolysis of acetylated H4 yields long peptidic HDAC substrates. HATs = histone acetyltransferases. (b) Solid-phase peptide synthesis (SPPS) using a radiolabeled acetyllysine building block (inset) yields a hepta-peptide that is not an HDAC substrate. (c) A long synthetic peptide bearing two distinctly radio-labeled acetyl groups illustrates promiscuity in HDAC activity.
