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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

2007; American Chemical Society; Volume: 6; Issue: 8 Linguagem: Inglês

10.1021/pr0701963

1535-3907

Mónica Carrera, Benito Cañas, Carmen Piñeiro, Jesús Vázquez, José M. Gallardo,

MXene and MAX Phase Materials

The 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. Keywords: proteomics • nucleoside diphosphate kinase • fish • de novo sequencing • mass spectrometry • selective ion reaction monitoring • NDK • SIRM