Differentiation between peptides containing acetylated or tri-methylated lysines by mass spectrometry: An application for determining lysine 9 acetylation and methylation of histone H3

Histone acetylation and methylation play a critical role in transcription and gene regulation. Identification of sites of lysine acetylation and methylation in histones or other proteins by mass spectrometry (MS) is of increasing interest. In this paper, we report the use of MS to differentiate betw...

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Bibliographic Details
Published in:Proteomics (Weinheim) 2004-01, Vol.4 (1), p.1-10
Main Authors: Zhang, Kangling, Yau, Peter M., Chandrasekhar, Bhaskar, New, Ron, Kondrat, Richard, Imai, Brian S., Bradbury, Morton E.
Format: Article
Language:English
Subjects:
Acetylation
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Fundamental and applied biological sciences. Psychology
Histone acetylation
Histone methylation
Histones
Histones - chemistry
Histones - metabolism
Lysine - analogs & derivatives
Lysine - chemistry
Lysine - classification
Mass spectrometry
Methylation
Miscellaneous
Peptides - analysis
Peptides - chemistry
Peptides - classification
Proteins
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
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Summary:Histone acetylation and methylation play a critical role in transcription and gene regulation. Identification of sites of lysine acetylation and methylation in histones or other proteins by mass spectrometry (MS) is of increasing interest. In this paper, we report the use of MS to differentiate between peptides containing acetylated or tri‐methylated lysines. High accuracy matrix‐assisted laser desorption/ionization‐time of flight MS gives better than five parts per million measurement accuracy, which is sufficient to verify acetylation and/or methylation. Electrospray ionization tandem mass spectrometry was used to assign modification sites and to differentiate acetylation from methylation. Typically, an immonium ion at m/z 98 corresponds to a mono‐methylated lysine and an immonium ion at m/z 126 corresponds to an acetylated lysine. The neutral loss ion (MH+‐59) is unique for a tri‐methylated lysine. For a peptide with two or more modification sites of acetylation or tri‐methylation or one site containing partial acetylation and tri‐methylation, the a2‐, b2‐type ion is the characteristic index for an acetylated lysine whereas the b2‐59 ion is indicative of a tri‐methylated lysine in the N‐terminus. The y‐type ions and y‐59 ions are charateristic of an acetylated lysine and a tri‐methylated lysine at the C‐terminus, respectively. We demonstrated that a lysine in a peptide modified by methylation or acetylation can be differentiated by MS using our method. Even if more then one lysine is present in a peptide and different modifications of this amino acid occur, they can be distinguished. This method was successful for the determination of the acetylation and methylation status of lysine 9 of histone H3 in chicken erythrocytes and human HeLa cell lines.
ISSN:1615-9853
1615-9861
DOI:10.1002/pmic.200300503