1. Imaging mass spectrometry using peptide isoelectric focusing
- Author
-
Ali R. Vaezzadeh, Jovan Simicevic, Catherine G. Zimmermann-Ivol, Alexis Chauvet, Patrice Francois, Denis F. Hochstrasser, Pierre Lescuyer, and Jacques P. M. Deshusses
- Subjects
Proteomics ,Staphylococcus aureus ,Analytical chemistry ,Peptide ,Mass spectrometry ,Peptide Mapping ,Mass spectrometry imaging ,Analytical Chemistry ,Species Specificity ,Tandem Mass Spectrometry/instrumentation/methods ,Tandem Mass Spectrometry ,ddc:576 ,Shotgun proteomics ,Spectroscopy ,chemistry.chemical_classification ,Chromatography ,Chemistry ,Isoelectric focusing ,Organic Chemistry ,Membrane Proteins/chemistry ,Membrane Proteins ,Staphylococcus aureus/chemistry/classification ,Membrane ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Proteomics/methods ,Membrane proteome ,Isoelectric Focusing ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/instrumentation/methods - Abstract
Imaging Mass Spectrometry (IMS) has emerged as a powerful technique in the field of proteomics. The use of Immobilized pH Gradient-IsoElectric Focusing (IPG-IEF) is also a new trend, as the first dimension of separation, in shotgun proteomics. We report a combination of these two outstanding technologies. This approach is based on the separation of shotgun-produced peptides by IPG-IEF. The peptides are then transferred by capillarity to a capture membrane, which is then scanned by the mass spectrometer to generate MS images. This high-throughput methodology allows a preview of the sample to be obtained in a single day. We report the application of this new pipeline for differential comparison of the membrane proteome of two different strains of Staphylococcus aureus bacteria in a proof-of-principle experiment.
- Published
- 2008