Analysis of Protein Tyrosine Phosphorylation by Nanoelectrospray Ionization High-Resolution Tandem Mass Spectrometry and Tyrosine-Targeted Product Ion Scanning
- 1 May 2003
- journal article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 75 (11) , 2724-2729
- https://doi.org/10.1021/ac020657y
Abstract
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.Keywords
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