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

Sweet, Steve M. M and Mardakheh, Faraz Khosravi and Ryan, Kevin J. P and Langton, Amy J and Heath, John K and Cooper, Helen J. (2008) Targeted online liquid chromatography electron capture dissociation mass spectrometry for the localization of sites of in vivo phosphorylation in human Sprouty2. Analytical Chemistry, 80 (17). pp. 6650-6657. ISSN 0003-2700

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URL of Published Version: http://pubs.acs.org/doi/abs/10.1021/ac800963a

Identification Number/DOI: 10.1021/ac800963a

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.

Type of Work:Article
Date:07 August 2008 (Publication)
School/Faculty:Schools (1998 to 2008) > School of Biosciences
Department:Structural Biology
Subjects:Q Science (General)
Institution:University of Birmingham
Copyright Holders:American Chemical Society
ID Code:123
Refereed:YES
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