High-sensitivity analysis of specific peptides in complex samples by selected MS/MS ion monitoring and linear ion trap mass spectrometry: application to biological studies
| Autor: | Anabel Marina, Santiago Lamas, Juan Miguel Redondo, Inmaculada Ortega-Pérez, Elisabet Miró Casas, Benito Cañas, David Garcia-Dorado, Antonio Martínez-Ruiz, Mónica Carrera, Horacio Serrano, José Manuel Gallardo, Pablo Bayón Martínez, Felipe Were, Daniel Lopez-Ferrer, Margarita Villar, Jesús Vázquez, Inmaculada Jorge |
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| Přispěvatelé: | Instituto de Salud Carlos III, Ministerio de Sanidad y Consumo (España), Fundación Ramón Areces |
| Rok vydání: | 2007 |
| Předmět: |
Proteomics
Tandem mass spectrometry Context (language use) Peptide Mass spectrometry p38 Mitogen-Activated Protein Kinases Mice Species Specificity Tandem Mass Spectrometry Fish Products Animals Humans Trypsin Cysteine HSP90 Heat-Shock Proteins Quadrupole ion trap Phosphorylation Spectroscopy chemistry.chemical_classification Linear ion trap Chromatography S-Nitrosothiols NFATC Transcription Factors Chemistry Cell Membrane Proteolytic enzymes Endothelial Cells Proteins Peptide Fragments Rats Gadiformes Connexin 43 Ion monitoring Electrophoresis Polyacrylamide Gel Ion trap Protein identification Protein Processing Post-Translational Post-translational modifications |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname |
| ISSN: | 1076-5174 |
| Popis: | 13 pages, 4 figures.-- PMID: 17960563 [PubMed].-- Printed version published Nov 2007.-- Inmaculada Jorge ... et al. Mass spectrometry (MS) is a technique of paramount importance in Proteomics, and developments in this field have been possible owing to novel MS instrumentation, experimental strategies, and bioinformatics tools. Today it is possible to identify and determine relative expression levels of thousands of proteins in a biological system by MS analysis of peptides produced by proteolytic digestion. In some situations, however, the precise characterization of a particular peptide species in a very complex peptide mixture is needed. While single-fragment ion-based scanning modes such as selected ion reaction monitoring (SIRM) or consecutive reaction monitoring (CRM) may be highly sensitive, they do not produce MS/MS information and their actual specificity must be determined in advance, a prerequisite that is not usually met in a basic research context. In such cases, the MS detector may be programmed to perform continuous MS/MS spectra on the peptide ion of interest in order to obtain structural information. This selected MS/MS ion monitoring (SMIM) mode has a number of advantages that are fully exploited by MS detectors that, like the linear ion trap, are characterized by high scanning speeds. In this work, we show some applications of this technique in the context of biological studies. These results were obtained by selecting an appropriate combination of scans according to the purpose of each one of these research scenarios. They include highly specific identification of proteins present in low amounts, characterization and relative quantification of post-translational modifications such as phosphorylation and S-nitrosylation and species-specific peptide identification. This work was supported by grants BIO2003-01805, BIO2006-10085, GR/SAL/0141/2004 (CAM), CAM BIO/0194/2006, the Fondo de Investigaciones Sanitarias (Ministerio de Sanidad y Consumo, Instituto Salud Carlos III, RECAVA), and by an institutional grant by Fundación Ramón Areces to CBMSO. |
| Databáze: | OpenAIRE |
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