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De novo mass spectrometry sequencing and characterization of species-specific peptides from nucleoside diphosphate kinase B for the classification of commercial fish species belonging to the family Merlucciidae

AuthorsCarrera, Mónica ; Cañas, Benito; Piñeiro, Carmen ; Vázquez, Jesús ; Gallardo, José Manuel
Nucleoside diphosphate Kinase
De novo sequencing
Mass spectrometry
Selective ion reaction monitoring
Issue Date10-Jul-2007
PublisherAmerican Chemical Society
CitationJournal of Proteome Research 6(8): 3070-3080 (2007)
AbstractThe characterization by de novo peptide sequencing of the different protein nucleoside diphosphate kinase B (NDK B) from all the commercial hakes and grenadiers belonging to the family Merlucciidae is reported. A classical proteomics approach, consisting of two-dimmensional gel electrophoresis, tryptic in-gel digestion of the excised spots, MALDI-TOF MS, LC−MS/MS, and nanoESI−MS/MS analyses, was followed for the purification and characterization of the different isoforms of the NDK B. Fragmentation spectra were used for de novo peptide sequence. A high degree of homology was found between the sequences of all the species studied and the NDK B sequence from Gillichthys mirabilis, which is accessible in the protein databases. Particular attention was paid to the differential characterization of species-specific peptides that could be used for fish authentication purposes. These findings allowed us to propose a rapid and effective classification method, based in the detection of these biomarker peptides using the selective ion reaction monitoring (SIRM) scan mode in mass spectrometry.
Description11 pages, 7 figures.-- PMID: 17622168 [PubMed].-- Supplementary information (Results and discussion, figure S1, table S1, 2 pages) available at: http://pubs.acs.org/doi/suppl/10.1021/pr0701963/suppl_file/pr0701963si20070521_060524.pdf.
Printed version published on Aug 3, 2007.
Publisher version (URL)http://dx.doi.org/10.1021/pr0701963
Appears in Collections:(IIM) Artículos
(CBM) Artículos
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