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Histone Acetylation and Deacetylation
The acetylation isoforms of histone H4 from butyrate-treated HeLa cells were separated by C 4 reverse-phase high pressure liquid chromatography and by polyacrylamide gel electrophoresis. Histone H4 bands were excised and digested in-gel with the endoprotease trypsin. Matrix-assisted laser desorption...
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Published in: | Molecular & cellular proteomics 2002-07, Vol.1 (7), p.500-508 |
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Main Authors: | , , , , , , , , |
Format: | Article |
Language: | English |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | The acetylation isoforms of histone H4 from butyrate-treated HeLa cells were separated by C 4 reverse-phase high pressure liquid chromatography and by polyacrylamide gel electrophoresis. Histone H4 bands were excised
and digested in-gel with the endoprotease trypsin. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry
was used to characterize the level of acetylation, and nanoelectrospray tandem mass spectrometric analysis of the acetylated
peptides was used to determine the exact sites of acetylation. Although there are 15 acetylation sites possible, only four
acetylated peptide sequences were actually observed. The tetra-acetylated form is modified at lysines 5, 8, 12, and 16, the
tri-acetylated form is modified at lysines 8, 12, and 16, and the di-acetylated form is modified at lysines 12 and 16. The
only significant amount of the mono-acetylated form was found at position 16. These results are consistent with the hypothesis
of a âzipâ model whereby acetylation of histone H4 proceeds in the direction of from Lys-16 to Lys-5, and deacetylation proceeds
in the reverse direction. Histone acetylation and deacetylation are coordinated processes leading to a non-random distribution
of isoforms. Our results also revealed that lysine 20 is di-methylated in all modified isoforms, as well as the non-acetylated
isoform of H4. |
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ISSN: | 1535-9476 1535-9484 |
DOI: | 10.1074/mcp.M200031-MCP200 |