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Electron capture dissociation mass spectrometric analysis of lysine-phosphorylated peptides
- Source :
- Bioscience Reports, Bioscience Reports, Portland Press, 2010, 30 (6), pp.433-443. ⟨10.1042/BSR20090167⟩
- Publication Year :
- 2010
- Publisher :
- Portland Press Ltd., 2010.
-
Abstract
- International audience; Phosphorylation of proteins is an essential signaling mechanism in eukaryotic and prokaryotic cells. Although N-phosphorylation of basic amino acid is known for its importance in biological systems, it is still poorly explored in terms of products and mechanisms. In this study, two mass spectrometry fragmentation methods, electron-capture dissociation (ECD) and collision-induced dissociation (CID), were tested as tools for analysis of N-phosphorylation of three model peptides, RKRSRAE, RKRARKE, and PLSRTLSVAAKK. The peptides were phosphorylated by the reaction with monopotassium phosphoramidate. The results were confirmed by the 1H NMR and 31P NMR studies. The ECD method was found useful for the localization of phosphorylation sites in unstable lysine-phosphorylated peptides. Its main advantage is a significant reduction of the neutral losses related to phosphoramidate moiety. Moreover, the results indicate that the ECD mass spectrometry may be useful for analysis of regioselectivity of the N-phosphorylation reaction. Stabilities of the obtained lysine-phosphorylated peptides under various conditions were also tested.
- Subjects :
- Phosphopeptides
phospholysine
inorganic chemicals
ECD, electron capture dissociation
CID, collision-induced dissociation
Lysine
Biophysics
Electrons
macromolecular substances
Mass spectrometry
S2
environment and public health
01 natural sciences
Biochemistry
Mass Spectrometry
Dissociation (chemistry)
ETD, electron transfer dissociation
03 medical and health sciences
ESI–MS, electrospray ionization MS
Fragmentation (mass spectrometry)
HMBC, heteronuclear multiple bond correlation
Phosphoric Acids
Amino Acid Sequence
Molecular Biology
Peptide sequence
030304 developmental biology
chemistry.chemical_classification
Original Paper
0303 health sciences
Electron-capture dissociation
phosphorylation
Chemistry
010401 analytical chemistry
Life Sciences
Phosphoramidate
Cell Biology
Amides
Magnetic Resonance Imaging
electron capture dissociation (ECD)
0104 chemical sciences
Amino acid
enzymes and coenzymes (carbohydrates)
post-translational modification
PKG, protein kinase G
bacteria
Peptides
Protein Processing, Post-Translational
Subjects
Details
- ISSN :
- 15734935 and 01448463
- Volume :
- 30
- Database :
- OpenAIRE
- Journal :
- Bioscience Reports
- Accession number :
- edsair.doi.dedup.....4d1f0644896a27f8e317aca91a61efeb