TY  - THES
T1  - Two-dimensional reversed-phase liquid chromatography coupled to MALDI TOF/TOF mass spectrometry : an approach for shotgun proteome analysis
A1  - Lasaosa,Maria
Y1  - 2008/12/04
N2  - Large-scale proteome analysis of complex biological mixtures requires high resolving power separations, high-throughput mass spectrometry and accurate detection, and bioinformatics tools. In the present work LC-MALDI TOF/TOF mass spectrometry was combined for the first time with a two-dimensional separation based on reversed-phase chromatography at high and low pH for the analysis of the cytosolic proteome of Corynebacterium glutamicum. The proteome coverage achieved by this approach, 55%, is the highest reported up to day for this bacterium. A total of 1644 proteins including single-peptide based identifications were identified. 
The sample was also analysed by other conventional methods in the proteome analysis. The classical 2D-PAGE approach presents more limitations and delivered 166 different proteins including enzymes of the main metabolic pathways. Complementary results were found at peptide level for the LC-MALDI and 2D-PAGE approaches. Fractions collected during the first dimension at high pH were analysed by LC-MALDI MS and compared with the analysis carried out by LC-ESI-IT MS, which identified 745 proteins [1]. Further comparison with results found in literature confirmed that the two-dimensional reversed phase combination with LC-MALDI MS/MS is a promising tool in the proteome analysis. The MALDI based approach showed higher sensitivity than other approaches and was able to identify proteins over a larger dynamic range, based on the codon adaptation indexes of the proteins.
KW  - LC-MS
KW  - MALDI-MS
KW  - Zweidimensionale Elektrophorese
KW  - Proteom
KW  - Proteomanalyse
KW  - Proteine
KW  - Peptide
CY  - Saarbrücken
PB  - Universitäts- und Landesbibliothek
AD  - Postfach 151141, 66041 Saarbrücken
UR  - http://scidok.sulb.uni-saarland.de/volltexte/2008/1806
ER  -