High accuracy mass spectrometry in large-scale analysis of protein phosphorylation.
With the appearance of a new generation of high-performance hybrid mass spectrometers, high accuracy (sub-parts-per-million) mass spectrometry is becoming increasingly available to a wider scientific community. Here we discuss the advantages of such mass spectrometric instrumentation in the global analysis of protein phosphorylation. We describe a detailed workflow for fractionation and enrichment of phosphopeptides from digests of whole cell/tissue lysates by strong cation exchange and TiO(2) chromatography and their subsequent measurement on an LTQ-Orbitrap mass spectrometer under several acquisition regimes.
Olsen
Jesper V
JV
Department of Proteomics and Signal Transduction, Max Planck Institute for Biochemistry, Martinsried, Germany.
Macek
Boris
B
D000595:Amino Acid Sequence; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D010748:Phosphopeptides / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D010750:Phosphoproteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D040901:Proteomics; D012680:Sensitivity and Specificity; D014025:Titanium / Q000737:chemistry; D014357:Trypsin / Q000378:metabolism
D010748:Phosphopeptides; D010750:Phosphoproteins; C009495:titanium dioxide; D014025:Titanium; D014357:Trypsin