Figure 3.

Pure HDAC1-F deacetylates histones in the absence of protein cofactors. (A) HDAC1-F was purified by using the protocol shown and the final step FLAG-peptide elution was examined on a 10% SDS/PAGE reducing gel, stained with Coomassie blue. Relative molecular mass is indicated in kilodaltons. (B) [³H]Acetate-labeled histones from butyrate-treated HeLa cells were purified as individual histones (or an H2A/H2B mixture) by electroelution, and a sample from each was run on an SDS/18% polyacrylamide gel and stained with Coomassie blue. Total acid extracted HeLa cellular histones were loaded in the far right lane as a reference and the positions of the individual histones are indicated (Right). (C) Pure HDAC1-F (25 ng) was incubated with the indicated histones (0.5 μg) and the average deacetylation for a given substrate was divided by the total dpm and calculated as the percent deacetylation.