title: Identification and quantitation of signal molecule-dependent protein phosphorylation.
abstract: Phosphoproteomics is a fast-growing field that aims at characterizing phosphorylated proteins in a cell or a tissue at a given time. Phosphorylation of proteins is an important regulatory mechanism in many cellular processes. Gel-free phosphoproteome technique involving enrichment of phosphopeptide coupled with mass spectrometry has proven to be invaluable to detect and characterize phosphorylated proteins. In this chapter, a gel-free quantitative approach involving (15)N metabolic labelling in combination with phosphopeptide enrichment by titanium dioxide (TiO2) and their identification by MS is described. This workflow can be used to gain insights into the role of signalling molecules such as cyclic nucleotides on regulatory networks through the identification and quantification of responsive phospho(proteins).
authors.affiliation: Department of Biochemistry, Cambridge Centre for Proteomics, Cambridge Systems Biology Centre, University of Cambridge, Cambridge, UK.
authors.forename: Arnoud
authors.initials: A
authors.lastname: Groen
authors.forename: Ludivine
authors.initials: L
authors.lastname: Thomas
authors.forename: Kathryn
authors.initials: K
authors.lastname: Lilley
authors.forename: Claudius
authors.initials: C
authors.lastname: Marondedze
mesh_terms: D002411:Cation Exchange Resins; D005591:Chemical Fractionation; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid; D003902:Detergents; D013058:Mass Spectrometry; D010748:Phosphopeptides / Q000737:chemistry / Q000378:metabolism; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D020543:Proteome / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction; D014025:Titanium
chemical_list: D002411:Cation Exchange Resins; D003902:Detergents; D010748:Phosphopeptides; D010750:Phosphoproteins; D020543:Proteome; C009495:titanium dioxide; D014025:Titanium