Study of acetylation on Ser/Thr/Tyr/Lys, and trimethylation on Lys using electrospray tandem mass spectrometry

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Abstract

Post-translational modifications (PTM) corresponding to a gain in mass of 42 Da are of increasing interest. It has been widely recognized that acetylation and trimethylation on Lys regulates gene transcription and silencing. In addition, it was recently discovered that acetylation of Ser and Thr residues on a signaling kinase can block its activation. In this paper, three series of model peptides were chemically synthesized to generate comparative MS data. Electrospray collision-induced dissociation tandem mass spectrometry was used to characterize the fragmentation pattern of acetylation on Ser, Thr, and Tyr residues. In separate experiments, the fragmentation pattern and efficiency were studied for acetylation and trimethylation on Lys. Our results confirmed those previously reported, that a characteristic immonium ion at m/z 126 corresponds to an acetylated Lys, and we further differentiated acetylation from trimethylation by their effects on peptide fragmentation efficiency. With the same primary sequence, a trimethylated peptide requires higher energy to fragment compared to the acetylated analogue. For peptides containing acetylated Ser, the y-60 and b-60 ions are commonly observed when the acetylation site is at, or close to, the C-terminus or N-terminus of the daughter ion, respectively; for acetylated Thr, in addition to y-60 and b-60 ions, y-42 ions are usually dominant. The loss of 42 Da and 60 Da can correspond to the loss of CH2CO through deacetylation and CH3COOH through β-elimination, respectively. Meanwhile, loss of 42 Da and 18 Da individually can also contribute to the loss of 60 Da. When peptide containing acetylated Tyr/Lys is fragmented, the acetyl group remains attached to their respective side-chains. The fragmentation pattern was similar whether the acetylation site was close to C-terminus or N-terminus of the peptide. This study provides a better understanding of the MSMS fragmentation character of peptides with acetylation on Ser, Thr, and Tyr, and the differences between acetylated and trimethylated Lys.

Original languageEnglish (US)
Pages (from-to)24-30
Number of pages7
JournalInternational Journal of Mass Spectrometry
Volume281
Issue number1-2
DOIs
StatePublished - Mar 15 2009

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acetylation
Acetylation
Mass spectrometry
Peptides
mass spectroscopy
peptides
fragmentation
Ions
ions
Transcription
genes
elimination
Phosphotransferases
Genes
Chemical activation
fragments
activation
dissociation
analogs
collisions

Keywords

  • Acetylation
  • Fragmentation
  • Fragmentation efficiency
  • Post-translational modification
  • Trimethylation

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Spectroscopy
  • Condensed Matter Physics
  • Instrumentation

Cite this

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title = "Study of acetylation on Ser/Thr/Tyr/Lys, and trimethylation on Lys using electrospray tandem mass spectrometry",
abstract = "Post-translational modifications (PTM) corresponding to a gain in mass of 42 Da are of increasing interest. It has been widely recognized that acetylation and trimethylation on Lys regulates gene transcription and silencing. In addition, it was recently discovered that acetylation of Ser and Thr residues on a signaling kinase can block its activation. In this paper, three series of model peptides were chemically synthesized to generate comparative MS data. Electrospray collision-induced dissociation tandem mass spectrometry was used to characterize the fragmentation pattern of acetylation on Ser, Thr, and Tyr residues. In separate experiments, the fragmentation pattern and efficiency were studied for acetylation and trimethylation on Lys. Our results confirmed those previously reported, that a characteristic immonium ion at m/z 126 corresponds to an acetylated Lys, and we further differentiated acetylation from trimethylation by their effects on peptide fragmentation efficiency. With the same primary sequence, a trimethylated peptide requires higher energy to fragment compared to the acetylated analogue. For peptides containing acetylated Ser, the y-60 and b-60 ions are commonly observed when the acetylation site is at, or close to, the C-terminus or N-terminus of the daughter ion, respectively; for acetylated Thr, in addition to y-60 and b-60 ions, y-42 ions are usually dominant. The loss of 42 Da and 60 Da can correspond to the loss of CH2CO through deacetylation and CH3COOH through β-elimination, respectively. Meanwhile, loss of 42 Da and 18 Da individually can also contribute to the loss of 60 Da. When peptide containing acetylated Tyr/Lys is fragmented, the acetyl group remains attached to their respective side-chains. The fragmentation pattern was similar whether the acetylation site was close to C-terminus or N-terminus of the peptide. This study provides a better understanding of the MSMS fragmentation character of peptides with acetylation on Ser, Thr, and Tyr, and the differences between acetylated and trimethylated Lys.",
keywords = "Acetylation, Fragmentation, Fragmentation efficiency, Post-translational modification, Trimethylation",
author = "Yan Li and Ball, {Haydn L.}",
year = "2009",
month = "3",
day = "15",
doi = "10.1016/j.ijms.2008.11.014",
language = "English (US)",
volume = "281",
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journal = "International Journal of Mass Spectrometry",
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TY - JOUR

T1 - Study of acetylation on Ser/Thr/Tyr/Lys, and trimethylation on Lys using electrospray tandem mass spectrometry

AU - Li, Yan

AU - Ball, Haydn L.

PY - 2009/3/15

Y1 - 2009/3/15

N2 - Post-translational modifications (PTM) corresponding to a gain in mass of 42 Da are of increasing interest. It has been widely recognized that acetylation and trimethylation on Lys regulates gene transcription and silencing. In addition, it was recently discovered that acetylation of Ser and Thr residues on a signaling kinase can block its activation. In this paper, three series of model peptides were chemically synthesized to generate comparative MS data. Electrospray collision-induced dissociation tandem mass spectrometry was used to characterize the fragmentation pattern of acetylation on Ser, Thr, and Tyr residues. In separate experiments, the fragmentation pattern and efficiency were studied for acetylation and trimethylation on Lys. Our results confirmed those previously reported, that a characteristic immonium ion at m/z 126 corresponds to an acetylated Lys, and we further differentiated acetylation from trimethylation by their effects on peptide fragmentation efficiency. With the same primary sequence, a trimethylated peptide requires higher energy to fragment compared to the acetylated analogue. For peptides containing acetylated Ser, the y-60 and b-60 ions are commonly observed when the acetylation site is at, or close to, the C-terminus or N-terminus of the daughter ion, respectively; for acetylated Thr, in addition to y-60 and b-60 ions, y-42 ions are usually dominant. The loss of 42 Da and 60 Da can correspond to the loss of CH2CO through deacetylation and CH3COOH through β-elimination, respectively. Meanwhile, loss of 42 Da and 18 Da individually can also contribute to the loss of 60 Da. When peptide containing acetylated Tyr/Lys is fragmented, the acetyl group remains attached to their respective side-chains. The fragmentation pattern was similar whether the acetylation site was close to C-terminus or N-terminus of the peptide. This study provides a better understanding of the MSMS fragmentation character of peptides with acetylation on Ser, Thr, and Tyr, and the differences between acetylated and trimethylated Lys.

AB - Post-translational modifications (PTM) corresponding to a gain in mass of 42 Da are of increasing interest. It has been widely recognized that acetylation and trimethylation on Lys regulates gene transcription and silencing. In addition, it was recently discovered that acetylation of Ser and Thr residues on a signaling kinase can block its activation. In this paper, three series of model peptides were chemically synthesized to generate comparative MS data. Electrospray collision-induced dissociation tandem mass spectrometry was used to characterize the fragmentation pattern of acetylation on Ser, Thr, and Tyr residues. In separate experiments, the fragmentation pattern and efficiency were studied for acetylation and trimethylation on Lys. Our results confirmed those previously reported, that a characteristic immonium ion at m/z 126 corresponds to an acetylated Lys, and we further differentiated acetylation from trimethylation by their effects on peptide fragmentation efficiency. With the same primary sequence, a trimethylated peptide requires higher energy to fragment compared to the acetylated analogue. For peptides containing acetylated Ser, the y-60 and b-60 ions are commonly observed when the acetylation site is at, or close to, the C-terminus or N-terminus of the daughter ion, respectively; for acetylated Thr, in addition to y-60 and b-60 ions, y-42 ions are usually dominant. The loss of 42 Da and 60 Da can correspond to the loss of CH2CO through deacetylation and CH3COOH through β-elimination, respectively. Meanwhile, loss of 42 Da and 18 Da individually can also contribute to the loss of 60 Da. When peptide containing acetylated Tyr/Lys is fragmented, the acetyl group remains attached to their respective side-chains. The fragmentation pattern was similar whether the acetylation site was close to C-terminus or N-terminus of the peptide. This study provides a better understanding of the MSMS fragmentation character of peptides with acetylation on Ser, Thr, and Tyr, and the differences between acetylated and trimethylated Lys.

KW - Acetylation

KW - Fragmentation

KW - Fragmentation efficiency

KW - Post-translational modification

KW - Trimethylation

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