Targeted online liquid chromatography electron capture dissociation mass spectrometry for the localization of sites of in vivo phosphorylation in human Sprouty2

Steve Sweet, FK Mardekheh, Kevin Ryan, AJ Langton, John Heath, Helen Cooper

Research output: Contribution to journalArticlepeer-review

23 Citations (Scopus)
121 Downloads (Pure)

Abstract

We demonstrate a strategy employing collision-induced dissociation for phosphopeptide discovery, followed by targeted electron capture dissociation (ECD) for site localization. The high mass accuracy and low background noise of the ECD mass spectra allow facile sequencing of coeluting isobaric phosphopeptides, with up to two isobaric phosphopeptides sequenced from a single mass spectrum. In contrast to the previously described neutral loss of dependent ECD method, targeted ECD allows analysis of both phosphotyrosine peptides and lower abundance phosphopeptides. The approach was applied to phosphorylation analysis of human Sprouty2, a regulator of receptor tyrosine kinase signaling. Fifteen sites of phosphorylation were identified, 11 of which are novel.
Original languageEnglish
Pages (from-to)6650-6657
Number of pages8
JournalAnalytical Chemistry
Volume80
Issue number17
DOIs
Publication statusPublished - 1 Jan 2008

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