Imaging mass spectrometry using peptide isoelectric focusing.
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.
Vaezzadeh
Ali R
AR
Biomedical Proteomics Research Group, Department of Structural Biology and Bioinformatics, Geneva University, 1 rue Michel Servet, 1211 Geneva, Switzerland. alireza.vaezzadeh@medecine.unige.ch
Simicevic
Jovan
J
Chauvet
Alexis
A
François
Patrice
P
Zimmermann-Ivol
Catherine G
CG
Lescuyer
Pierre
P
Deshusses
Jacques P M
JP
Hochstrasser
Denis F
DF
D007525:Isoelectric Focusing; D008565:Membrane Proteins / Q000737:chemistry*; D010449:Peptide Mapping; D040901:Proteomics / Q000379:methods*; D013045:Species Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation* / Q000379:methods*; D013211:Staphylococcus aureus / Q000737:chemistry* / Q000145:classification; D053719:Tandem Mass Spectrometry / Q000295:instrumentation / Q000379:methods
D008565:Membrane Proteins