title: An initial characterization of the serum phosphoproteome.
abstract: Phosphorylation is a dynamic post-translational protein modification that is the basis of a general mechanism for maintaining and regulating protein structure and function, and of course underpins key cellular processes through signal transduction. In the last several years, many studies of large-scale profiling of phosphoproteins and mapping phosphorylation sites from cultured human cells or tissues by mass spectrometry technique have been published; however, there is little information on general (or global) phosphoproteomic characterization and description of the content of phosphoprotein analytes within the circulation. Circulating phosphoproteins and phosphopeptides could represent important disease biomarkers because of their well-known importance in cellular function, and these analytes frequently are mutated and activated in human diseases such as cancer. Here, we report an initial attempt to characterize the phosphoprotein content of serum. To accomplish this, we developed a method in which phosphopeptides are enriched from digested serum proteins and analyzed by LC-MS/MS using LTQ-Orbitrap (CID) and LTQ-ETD mass spectrometers. With this approach, we identified approximately 100 unique phosphopeptides with stringent filtering criteria and a lower than 1% false discovery rate.
authors.affiliation: Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia 20110, USA.
authors.forename: Weidong
authors.initials: W
authors.lastname: Zhou
authors.forename: Mark M
authors.initials: MM
authors.lastname: Ross
authors.forename: Alessandra
authors.initials: A
authors.lastname: Tessitore
authors.forename: David
authors.initials: D
authors.lastname: Ornstein
authors.forename: Amy
authors.initials: A
authors.lastname: Vanmeter
authors.forename: Lance A
authors.initials: LA
authors.lastname: Liotta
authors.forename: Emanuel F
authors.initials: EF
authors.lastname: Petricoin
mesh_terms: D001798:Blood Proteins / Q000032:analysis*; D002853:Chromatography, Liquid; D006801:Humans; D008722:Methods; D010748:Phosphopeptides / Q000097:blood; D010750:Phosphoproteins / Q000097:blood*; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D001798:Blood Proteins; D010748:Phosphopeptides; D010750:Phosphoproteins; D020543:Proteome