TY - JOUR
T1 - Taking the shortcut for high-throughput shotgun proteomic analysis of bacteria
AU - Hartmann, Erica Marie
AU - Allain, François
AU - Gaillard, Jean Charles
AU - Pible, Olivier
AU - Armengaud, Jean
N1 - Publisher Copyright:
© Springer Science+Business Media New York 2014.
PY - 2014
Y1 - 2014
N2 - Currently, proteomic tools are able to establish a complete list of the most abundant proteins present in a sample, providing the opportunity to study at high resolution the physiology of any bacteria for which the genome sequence is available. For a comprehensive list, proteins should be first resolved into fractions that are then proteolyzed by trypsin. The resulting peptide mixtures are analyzed by a high-throughput tandem mass spectrometer that records thousands of MS/MS spectra for each fraction. These spectra are then assigned to peptides, which are used as evidence of the existence of proteins. In addition to generating a list of protein identifications, this shortcut to proteomics uses the number of spectra recorded for each protein to quantify the observations. Here, we describe one of the most simple sample preparation methods for high-throughput proteomics of bacteria, as well as the subsequent data processing to extract quantitative information based on the spectral count approach.
AB - Currently, proteomic tools are able to establish a complete list of the most abundant proteins present in a sample, providing the opportunity to study at high resolution the physiology of any bacteria for which the genome sequence is available. For a comprehensive list, proteins should be first resolved into fractions that are then proteolyzed by trypsin. The resulting peptide mixtures are analyzed by a high-throughput tandem mass spectrometer that records thousands of MS/MS spectra for each fraction. These spectra are then assigned to peptides, which are used as evidence of the existence of proteins. In addition to generating a list of protein identifications, this shortcut to proteomics uses the number of spectra recorded for each protein to quantify the observations. Here, we describe one of the most simple sample preparation methods for high-throughput proteomics of bacteria, as well as the subsequent data processing to extract quantitative information based on the spectral count approach.
KW - Bacterial proteome
KW - High-throughput proteomics
KW - Quantitative proteomics
KW - Spectral count
KW - Tandem mass spectrometry
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U2 - 10.1007/978-1-4939-1261-2_16
DO - 10.1007/978-1-4939-1261-2_16
M3 - Article
C2 - 25172287
AN - SCOPUS:84908252371
SN - 1064-3745
VL - 1197
SP - 275
EP - 285
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
ER -