Analysis of Protein Tyrosine Phosphorylation by Nanoelectrospray Ionization High-Resolution Tandem Mass Spectrometry and Tyrosine-Targeted Product Ion Scanning

A novel highly sensitive strategy is introduced for analysis of tyrosine phosphorylation in previously identified proteins channelling for this aim all analytical and sequence information available. Nanoelectrospray high-resolution MS/MS analysis is targeted to precalculated m/z values corresponding...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2003-06, Vol.75 (11), p.2724-2729
Main Authors: Salek, Mogjiborahman, Alonso, Angel, Pipkorn, R, Lehmann, Wolf D
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Analysis
Cell Line
ErbB Receptors - metabolism
Humans
Ions
Molecular Sequence Data
Nanotechnology
Peptide Fragments - analysis
Peptide Fragments - chemistry
Phosphates
Phosphoproteins - analysis
Phosphoproteins - metabolism
Phosphorylation
Phosphotyrosine - analysis
Phosphotyrosine - chemistry
Proteins
Spectrometry, Mass, Electrospray Ionization - methods
Trypsin - metabolism
Tyrosine - metabolism
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Summary:A novel highly sensitive strategy is introduced for analysis of tyrosine phosphorylation in previously identified proteins channelling for this aim all analytical and sequence information available. Nanoelectrospray high-resolution MS/MS analysis is targeted to precalculated m/z values corresponding to phosphotyrosine-containing tryptic peptides. Identification of these peptides is supported by the occurrence of the phosphotyrosine immonium ion at m/z 216, neutral loss of 79.97/z (= loss of HPO3), and similarity of the fragmentation patterns of phosphotyrosine-containing peptides with their nonphosphorylated analogues. This tyrosine-targeted tandem mass spectrometry strategy is demonstrated for epidermal growth factor receptor showing that phosphotyrosine-containing tryptic peptides invisible in the survey spectrum can be safely identified.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac020657y