Mapping of phosphorylation sites by LC-MS/MS.
Reversible protein phosphorylation ranks among the most important post-translational modifications that occurs in the cell. It is therefore highly relevant to elucidate the phosphorylation states of a given biological system, albeit challenging. Most notably the often low stoichiometry of phosphorylation is inherently incompatible with standard LC-MS analysis of a complex protein digest mixture, primarily due to the relative low dynamic range of current mass analyzers. Therefore a need for specific enrichment of phosphorylated peptides or proteins exists. Significant progress surrounding the biochemical analysis of reversible protein phosphorylation in the past years has led to the development of several new techniques to isolate or enrich phosphopeptides, particularly in large-scale analyses. This chapter deals with three such examples.
Gerrits
Bertran
B
Functional Genomics Center Zurich, University of Zurich and Swiss Federal Institute of Technology, Zurich, Switzerland.
Bodenmiller
Bernd
B
D053000:Analytic Sample Preparation Methods; D001665:Binding Sites; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid / Q000379:methods*; D030562:Databases, Protein; D050091:Dendrimers / Q000737:chemistry; D008863:Microspheres; D009943:Organophosphorus Compounds / Q000737:chemistry; D010446:Peptide Fragments / Q000032:analysis / Q000302:isolation & purification; D010750:Phosphoproteins / Q000737:chemistry; D010766:Phosphorylation; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014025:Titanium / Q000737:chemistry
D050091:Dendrimers; D009943:Organophosphorus Compounds; D010446:Peptide Fragments; D010750:Phosphoproteins; D011506:Proteins; C434331:phosphoramidite; C009495:titanium dioxide; D014025:Titanium