English   español  
Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/27498
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:


Extensive De Novo sequencing of new parvalbumin isoforms using a novel combination of bottom-up proteomics, accurate molecular mass measurement by FTICR-MS, and selected MS/MS Ion monitoring

AuthorsCarrera, Mónica ; Cañas, Benito; Vázquez, Jesús ; Gallardo, José Manuel
De novo sequencing
Selected MS/MS ion monitoring
Issue Date2010
PublisherAmerican Chemical Society
CitationJournal of Proteome Research 9: 4393–4406 (2010)
AbstractParvalbumins (PRVB) (11.20-11.55 kDa) are considered the major fish allergens. In this work, we propose a novel strategy for extensive characterization of this group of proteins based on the integration of a classical Bottom-Up proteomics approach with accurate Mr determination by FTICR-MS of intact proteins and selected MS/MS ion monitoring (SMIM) of peptide mass gaps. For each PRVB, mass spectra obtained by LC-ESI-IT-MS/MS from two digests (trypsin, Glu-C) were de novo sequenced manually with help of two programs (PEAKS, DeNovoX). The deduced peptide sequences were arranged and the theoretical Mr for the resulting sequences was calculated. Experimental Mr for each PRVB was measured with high mass accuracy by FTICR-MS (0.05-4.47 ppm). The masses of several missing peptide gaps were estimated by comparing the theoretical and experimentalMr, and the MS/MS spectra corresponding to these ions were obtained by LC-ESI-IT-MS/MS in the SMIM scanning mode. Finally, all peptide sequences were combined to generate the final protein sequences. This approach allowed the complete de novo MS-sequencing of 25 new PRVB isoforms. These new sequences belong to 11 different species from the Merlucciidae family, organisms for which genomes remain unsequenced. This study constitutes the report accounting for the higher number of new proteins completely sequenced making use of MS-based techniques only.
Publisher version (URL)http://dx.doi.org/10.1021/pr100163e
Appears in Collections:(IIM) Artículos
Files in This Item:
There are no files associated with this item.
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.