Charge state-selective separation of peptides by reversible modification of amino groups and strong cation-exchange chromatography: Evaluation in proteomic studies using peptide-centric database searches

Here we describe an integrated approach for the selective separation of peptides from complex mixtures using strong cation-exchange chromatography. The procedure exploits the charge differences produced by reversible modification of primary amino groups in peptides, enabling their separation into th...

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Bibliographic Details
Published in:Journal of proteomics 2011-09, Vol.74 (10), p.2210-2213
Main Authors: Betancourt, Lázaro H., Sánchez, Aniel, Pérez, Yasset, Fernandez de Cossio, Jorge, Gil, Jeovanis, Toledo, Patricia, Iguchi, Seiji, Aimoto, Saburo, González, Luis J., Padrón, Gabriel, Takao, Toshifumi, Besada, Vladimir
Format: Article
Language:English
Subjects:
Biological and medical sciences
Cation-exchange
Diverse techniques
Fundamental and applied biological sciences. Psychology
Molecular and cellular biology
Peptide-centric-database
Proteomics
Selective-peptide-isolation
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Summary:Here we describe an integrated approach for the selective separation of peptides from complex mixtures using strong cation-exchange chromatography. The procedure exploits the charge differences produced by reversible modification of primary amino groups in peptides, enabling their separation into three major fractions: 1) neutral peptides 2) peptides with one positive charge and 3) peptides with 2 or more positive charges. The procedure demonstrated an excellent selectivity which allowed restricted MS/MS ion searches with peptide-centric databases. [Display omitted] ► Complex mixture of tryptic peptides are exclusively separated by charge state. ► The high selectivity obtained improves confidence in the identification. ► Peptide centric databases searches are effectively used.
ISSN:1874-3919
DOI:10.1016/j.jprot.2011.04.029