Histone acetylation and deacetylation

identification of acetylation and methylation sites of HeLa histone H4 by mass spectrometry.

Kangling Zhang, Katherine E. Williams, Lan Huang, Peter Yau, Joseph S. Siino, E. Morton Bradbury, Patrick R. Jones, Michael J. Minch, Alma L. Burlingame

Research output: Contribution to journalArticle

130 Citations (Scopus)

Abstract

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 "zip" 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.

Original languageEnglish (US)
Pages (from-to)500-508
Number of pages9
JournalMolecular & cellular proteomics : MCP
Volume1
Issue number7
StatePublished - Jul 2002
Externally publishedYes

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Acetylation
Methylation
Histones
Mass spectrometry
Mass Spectrometry
Lysine
Protein Isoforms
High pressure liquid chromatography
Peptides
Butyrates
Reverse-Phase Chromatography
Electrophoresis
HeLa Cells
Trypsin
Ionization
Polyacrylamide Gel Electrophoresis
Desorption
Lasers
Gels
High Pressure Liquid Chromatography

ASJC Scopus subject areas

  • Biochemistry

Cite this

Zhang, K., Williams, K. E., Huang, L., Yau, P., Siino, J. S., Bradbury, E. M., ... Burlingame, A. L. (2002). Histone acetylation and deacetylation: identification of acetylation and methylation sites of HeLa histone H4 by mass spectrometry. Molecular & cellular proteomics : MCP, 1(7), 500-508.

Histone acetylation and deacetylation : identification of acetylation and methylation sites of HeLa histone H4 by mass spectrometry. / Zhang, Kangling; Williams, Katherine E.; Huang, Lan; Yau, Peter; Siino, Joseph S.; Bradbury, E. Morton; Jones, Patrick R.; Minch, Michael J.; Burlingame, Alma L.

In: Molecular & cellular proteomics : MCP, Vol. 1, No. 7, 07.2002, p. 500-508.

Research output: Contribution to journalArticle

Zhang, K, Williams, KE, Huang, L, Yau, P, Siino, JS, Bradbury, EM, Jones, PR, Minch, MJ & Burlingame, AL 2002, 'Histone acetylation and deacetylation: identification of acetylation and methylation sites of HeLa histone H4 by mass spectrometry.', Molecular & cellular proteomics : MCP, vol. 1, no. 7, pp. 500-508.
Zhang, Kangling ; Williams, Katherine E. ; Huang, Lan ; Yau, Peter ; Siino, Joseph S. ; Bradbury, E. Morton ; Jones, Patrick R. ; Minch, Michael J. ; Burlingame, Alma L. / Histone acetylation and deacetylation : identification of acetylation and methylation sites of HeLa histone H4 by mass spectrometry. In: Molecular & cellular proteomics : MCP. 2002 ; Vol. 1, No. 7. pp. 500-508.
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