title: Analytical methodologies for the detection and structural characterization of phosphorylated proteins.
abstract: Phosphorylation of proteins is a frequent post-translational modification affecting a great number of fundamental cellular functions in living organisms. Because of its key role in many biological processes, much effort has been spent over the time on the development of analytical methodologies for characterizing phosphoproteins. In the past decade, mass spectrometry-based techniques have emerged as a viable alternative to more traditional methods of phosphorylation analysis, providing accurate information for a purified protein on the number of the occurring phosphate groups and their exact localization on the polypeptide sequence. This review summarizes the analytical methodologies currently available for the analysis of protein phosphorylation, emphasizing novel mass spectrometry (MS) technologies and dedicated biochemical procedures that have been recently introduced in this field. A formidable armamentarium is now available for selective enrichment, exaustive structural characterization and quantitative determination of the modification degree for phosphopeptides/phosphoproteins. These methodologies are now successfully applied to the global analysis of cellular proteome repertoire according a holistic approach, allowing the quantitative study of phosphoproteomes on a dynamic time-course basis. The enormous complexity of the protein phosphorylation pattern inside the cell and its dynamic modification will grant important challenges to future scientists, contributing significantly to deeper insights into cellular processes and cell regulation.
authors.affiliation: Proteomics & Mass Spectrometry Laboratory, ISPAAM, National Research Council, via Argine 1085, 80147 Naples, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: D'Ambrosio
authors.forename: Anna Maria
authors.initials: AM
authors.lastname: Salzano
authors.forename: Simona
authors.initials: S
authors.lastname: Arena
authors.forename: Giovanni
authors.initials: G
authors.lastname: Renzone
authors.forename: Andrea
authors.initials: A
authors.lastname: Scaloni
mesh_terms: D015180:Electrophoresis, Gel, Two-Dimensional; D008956:Models, Chemical; D015394:Molecular Structure; D010750:Phosphoproteins / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D010750:Phosphoproteins