title: Clinical use of phosphorylated proteins in blood serum analysed by immobilised metal ion affinity chromatography and mass spectrometry.
abstract: The process of protein phosphorylation in cells is well studied in the context of a wide range of biologic functions such as signalling, cell cycle, cell growth and differentiation, and others. In contrast, little progress has been made in the investigation of protein phosphorylation specifically in blood. Here, we focussed on the phosphoproteome in human blood serum to study its extent and characteristics, and to explore the potential clinical utility. Immobilised metal ion affinity chromatography (IMAC) for the enrichment of intact phosphorylated proteins and label-free liquid chromatography-mass spectrometry (LC-MS(E)) were used for the molecular analysis of a large number of serum samples. To obtain high-confidence results, phosphorylated peptides had to be detected in at least 2 out of 3 technical replicates per sample and in >70% of the serum samples drawn from 80 volunteers. Individual analysis of these 80 non-pooled samples resulted in the detection of 5825 unique phosphorylated peptides after filtering, which corresponded to 502 unique proteins. The results provided evidence that blood serum may be an untapped source of phosphoproteins suitable for potential use in understanding disease pathophysiology and for identification of disease and drug response biomarkers. This article is part of a Special Issue entitled: Integrated omics.
authors.affiliation: Department of Chemical Engineering and Biotechnology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QT, United Kingdom. jj316@cam.ac.uk
authors.forename: Julian A J
authors.initials: JA
authors.lastname: Jaros
authors.forename: Paul C
authors.initials: PC
authors.lastname: Guest
authors.forename: Hassan
authors.initials: H
authors.lastname: Ramoune
authors.forename: Matthias
authors.initials: M
authors.lastname: Rothermundt
authors.forename: F Markus
authors.initials: FM
authors.lastname: Leweke
authors.forename: Daniel
authors.initials: D
authors.lastname: Martins-de-Souza
authors.forename: Sabine
authors.initials: S
authors.lastname: Bahn
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000032:analysis / Q000097:blood; D001798:Blood Proteins / Q000032:analysis / Q000378:metabolism*; D002846:Chromatography, Affinity / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D010750:Phosphoproteins / Q000032:analysis / Q000097:blood*
chemical_list: D015415:Biomarkers; D001798:Blood Proteins; D010750:Phosphoproteins