English   español  
Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/211306
Share/Impact:
Statistics
logo share SHARE logo core CORE   Add this article to your Mendeley library MendeleyBASE

Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL | DATACITE
Exportar a otros formatos:

Title

A combination of neutral loss and targeted product ion scanning with two enzymatic digestions facilitates the comprehensive mapping of phosphorylation sites

AuthorsCasado-Vela, Juan; Ruiz, Edgar J.; Nebreda, Ángel R.; Casal, J. Ignacio
KeywordsCdc25
Linear ion trap
Neutral loss
Phosphorylation analysis
TPIS
Issue Date23-Jul-2007
PublisherJohn Wiley & Sons
CitationProteomics 7: 2522-2529 (2007)
AbstractWe propose here a new strategy for the exhaustive mapping of phosphorylation sites in the Xenopus laevis Cdc25 phosphatase, which regulates cell cycle progression in eukaryotic cells. Two different MS analyses in a linear IT were used to identify the phosphorylated residues. First, a data‐dependent neutral loss (DDNL) analysis triggered the fragmentation of peptides that show enhanced neutral loss of phosphoric acid. Second, a targeted product ion scanning (TPIS) mass analysis was carried out in which MS2 events are triggered for specific m/z values. Full coverage of the protein sequence was obtained by combining the two analyses with two enzymatic digestions, trypsin and chymotrypsin, yielding a comprehensive map of the phosphorylation sites. Previous reports have shown Cdc25C to be phosphorylated by Cdc2–cyclin B at four residues (Thr48, Thr67, Thr138 and Ser205). By using this combination of scan modes, we have identified four additional phosphorylation sites (Thr86, Ser99, Thr112 and Ser163) in a recombinant Cdc25C protein containing 198 residues of the NH2‐terminal noncatalytic domain. The sensitivity of this combined approach makes it extremely useful for the comprehensive characterization of phosphorylation sites, virtually permitting complete coverage of the protein sequence with peptides within the mass detection range of the linear IT.
Description9 p-.5 fig.-2 tab.
Publisher version (URL)https://doi.org/10.1002/pmic.200700026
URIhttp://hdl.handle.net/10261/211306
DOI10.1002/pmic.200700026
ISSN1615-9853
E-ISSN1615-9861
Appears in Collections:(CIB) Artículos
Files in This Item:
File Description SizeFormat 
accesoRestringido.pdfRestringido15,38 kBAdobe PDFThumbnail
View/Open
Show full item record
Review this work
 

Related articles:


WARNING: Items in Digital.CSIC are protected by copyright, with all rights reserved, unless otherwise indicated.