title: Phosphoproteome analysis.
abstract: Protein phosphorylation is directly or indirectly involved in all important cellular events. The understanding of its regulatory role requires the discovery of the proteins involved in these processes and how, where and when protein phosphorylation takes place. Investigation of the phosphoproteome of a cell is becoming feasible today although it still represents a very difficult task especially if quantitative comparisons have to be made. Several different experimental strategies can be employed to explore phosphoproteomes and this review will cover the most important ones such as incorporation of radiolabeled phosphate into proteins, application of specific antibodies against phosphorylated residues and direct staining of phosphorylated proteins in polyacrylamide gels. Moreover, methods to enrich phosphorylated proteins such as affinity chromatography (IMAC) and immunoprecipitation as well as mass spectrometry for identification of phosphorylated peptides and phosphorylation sites are also described.
authors.affiliation: Sienabiotech S.p.A., Discovery Research, Siena, Italy. rraggiaschi@sienabiotech.com
authors.forename: Roberto
authors.initials: R
authors.lastname: Raggiaschi
authors.forename: Stefano
authors.initials: S
authors.lastname: Gotta
authors.forename: Georg C
authors.initials: GC
authors.lastname: Terstappen
mesh_terms: D000180:Acrylic Resins / Q000032:analysis; D002846:Chromatography, Affinity / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D047468:Immunoprecipitation / Q000379:methods; D007553:Isotope Labeling / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D010749:Phosphoprotein Phosphatases / Q000737:chemistry; D010750:Phosphoproteins / Q000032:analysis* / Q000737:chemistry; D010766:Phosphorylation; D020543:Proteome / Q000032:analysis*; D011869:Radioligand Assay / Q000379:methods
chemical_list: D000180:Acrylic Resins; D010750:Phosphoproteins; D020543:Proteome; C016680:polyacrylamide gels; D010749:Phosphoprotein Phosphatases