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

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• 作者：Mó ; nica Carrera ; Benito Cañ ; as ; Carmen Piñ ; eiro ; Jesú ; s Vá ; zquez ; Jos&eacute ; Manuel Gallardo
• 刊名：Journal of Proteome Research
• 出版年：2007
• 出版时间：August 2007
• 年：2007
• 卷：6
• 期：8
• 页码：3070 - 3080
• 全文大小：589K
• 年卷期：v.6,no.8(August 2007)
• ISSN：1535-3907

文摘

The characterization by de novo peptide sequencing of the different protein nucleoside diphosphatekinase B (NDK B) from all the commercial hakes and grenadiers belonging to the family Merlucciidaeis reported. A classical proteomics approach, consisting of two-dimmensional gel electrophoresis, trypticin-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 foundbetween 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 differentialcharacterization of species-specific peptides that could be used for fish authentication purposes. Thesefindings allowed us to propose a rapid and effective classification method, based in the detection ofthese biomarker peptides using the selective ion reaction monitoring (SIRM) scan mode in massspectrometry.