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Identification of single and double sites of phosphorylation by ECD FT-ICR/MS in peptides related to the phosphorylation site domain of the myristoylated alanine-rich C kinase protein

Woodling, Kellie A; Eyler, John R; Tsybin, Yury O; Nilsson, Carol L LU ; Marshall, Alan G; Edison, Arthur S; Al-Naggar, Iman M and Bubb, Michael R (2007) In Journal of the American Society for Mass Spectrometry 18(12). p.45-2137
Abstract

A series of phosphorylated test peptides was studied by electron capture dissociation Fourier transform ion cyclotron resonance mass spectrometry (ECD FT-ICR MS). The extensive ECD-induced fragmentation made identification of phosphorylation sites for these peptides straightforward. The site(s) of initial phosphorylation of a synthetic peptide with a sequence identical to that of the phosphorylation site domain (PSD) of the myristoylated alanine-rich C kinase (MARCKS) protein was then determined. Despite success in analyzing fragmentation of the smaller test peptides, a unique site on the PSD for the first step of phosphorylation could not be identified because the phosphorylation reaction produced a heterogeneous mixture of products.... (More)

A series of phosphorylated test peptides was studied by electron capture dissociation Fourier transform ion cyclotron resonance mass spectrometry (ECD FT-ICR MS). The extensive ECD-induced fragmentation made identification of phosphorylation sites for these peptides straightforward. The site(s) of initial phosphorylation of a synthetic peptide with a sequence identical to that of the phosphorylation site domain (PSD) of the myristoylated alanine-rich C kinase (MARCKS) protein was then determined. Despite success in analyzing fragmentation of the smaller test peptides, a unique site on the PSD for the first step of phosphorylation could not be identified because the phosphorylation reaction produced a heterogeneous mixture of products. Some molecules were phosphorylated on the serine closest to the N-terminus, and others on one of the two serines closest to the C-terminus of the peptide. Although no definitive evidence for phosphorylation on either of the remaining two serines in the PSD was found, modification there could not be ruled out by the ECD fragmentation data.

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publication status
published
keywords
Amino Acid Sequence, Intracellular Signaling Peptides and Proteins, Mass Spectrometry, Membrane Proteins, Molecular Sequence Data, Molecular Weight, Peptides, Phosphorylation, Protein Processing, Post-Translational, Protein Structure, Tertiary, Spectroscopy, Fourier Transform Infrared, Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.
in
Journal of the American Society for Mass Spectrometry
volume
18
issue
12
pages
9 pages
publisher
Elsevier
external identifiers
  • scopus:36349015923
ISSN
1044-0305
DOI
10.1016/j.jasms.2007.09.010
language
English
LU publication?
no
id
92dc5391-5553-4df1-bc03-40c4ab04a427
date added to LUP
2017-05-16 10:32:03
date last changed
2017-05-21 05:01:36
@article{92dc5391-5553-4df1-bc03-40c4ab04a427,
  abstract     = {<p>A series of phosphorylated test peptides was studied by electron capture dissociation Fourier transform ion cyclotron resonance mass spectrometry (ECD FT-ICR MS). The extensive ECD-induced fragmentation made identification of phosphorylation sites for these peptides straightforward. The site(s) of initial phosphorylation of a synthetic peptide with a sequence identical to that of the phosphorylation site domain (PSD) of the myristoylated alanine-rich C kinase (MARCKS) protein was then determined. Despite success in analyzing fragmentation of the smaller test peptides, a unique site on the PSD for the first step of phosphorylation could not be identified because the phosphorylation reaction produced a heterogeneous mixture of products. Some molecules were phosphorylated on the serine closest to the N-terminus, and others on one of the two serines closest to the C-terminus of the peptide. Although no definitive evidence for phosphorylation on either of the remaining two serines in the PSD was found, modification there could not be ruled out by the ECD fragmentation data.</p>},
  author       = {Woodling, Kellie A and Eyler, John R and Tsybin, Yury O and Nilsson, Carol L and Marshall, Alan G and Edison, Arthur S and Al-Naggar, Iman M and Bubb, Michael R},
  issn         = {1044-0305},
  keyword      = {Amino Acid Sequence,Intracellular Signaling Peptides and Proteins,Mass Spectrometry,Membrane Proteins,Molecular Sequence Data,Molecular Weight,Peptides,Phosphorylation,Protein Processing, Post-Translational,Protein Structure, Tertiary,Spectroscopy, Fourier Transform Infrared,Journal Article,Research Support, Non-U.S. Gov't,Research Support, U.S. Gov't, Non-P.H.S.},
  language     = {eng},
  number       = {12},
  pages        = {45--2137},
  publisher    = {Elsevier},
  series       = {Journal of the American Society for Mass Spectrometry},
  title        = {Identification of single and double sites of phosphorylation by ECD FT-ICR/MS in peptides related to the phosphorylation site domain of the myristoylated alanine-rich C kinase protein},
  url          = {http://dx.doi.org/10.1016/j.jasms.2007.09.010},
  volume       = {18},
  year         = {2007},
}