Use of gas-phase fractionation to increase protein identifications : application to the peroxisome.
Gas-phase fractionation (GPF), defined as iterative mass spectrometric interrogations of a sample over multiple smaller mass-to-charge (m/z) ranges, enables the ions selected for collision-induced dissociation to come from a greater number of unique peptides compared to the ions selected from the wide mass range scan in automated liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. GPF is described as a means to achieve higher proteome coverage than multiple LC-MS/MS analyses of unfractionated complex peptide mixtures. It is applied to organellar proteomics through analysis of yeast peroxisomal proteins obtained from a discontinuous Nycodenz gradient fraction known to be enriched with yeast peroxisomal membrane proteins.
Kennedy
Jacob
J
ZymoGenetics Inc., Seattle, WA, USA.
Yi
Eugene C
EC
D002197:Capillary Action; D002458:Cell Fractionation / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D005740:Gases; D013058:Mass Spectrometry / Q000379:methods; D015388:Organelles / Q000648:ultrastructure; D020675:Peroxisomes / Q000648:ultrastructure*; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
D005740:Gases; D011506:Proteins