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.
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.
Carrera
Mónica
M
Departamento Tecnología de Alimentos, Instituto de Investigaciones Marinas (IIM-CSIC), Vigo, Pontevedra, Spain. mcarrera@iim.csic.es
Cañas
Benito
B
Piñeiro
Carmen
C
Vázquez
Jesús
J
Gallardo
José Manuel
JM
D000595:Amino Acid Sequence; D000818:Animals; D029941:Fish Proteins / Q000032:analysis*; D005399:Fishes / Q000378:metabolism*; D008969:Molecular Sequence Data; D054778:NM23 Nucleoside Diphosphate Kinases / Q000032:analysis* / Q000737:chemistry; D010455:Peptides / Q000032:analysis*; D013045:Species Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D029941:Fish Proteins; D054778:NM23 Nucleoside Diphosphate Kinases; D010455:Peptides