title: Evaluation of phosphopeptide enrichment strategies for quantitative TMT analysis of complex network dynamics in cancer-associated cell signalling.
abstract: Defining alterations in signalling pathways in normal and malignant cells is becoming a major field in proteomics. A number of different approaches have been established to isolate, identify and quantify phosphorylated proteins and peptides. In the current report, a comparison between SCX prefractionationversusan antibody based approach, both coupled to TiO2enrichment and applied to TMT labelled cellular lysates, is described. The antibody strategy was more complete for enriching phosphopeptides and allowed the identification of a large set of proteins known to be phosphorylated (715 protein groups) with a minimum number of not previously known phosphorylated proteins (2).
authors.affiliation: Proteomics and Molecular Cell Dynamics, Center for Nephrology, School of Life and Medical Sciences, University College London, Royal Free Campus, Rowland Hill Street, London NW3 2PF, United Kingdom.
authors.forename: Benedetta
authors.initials: B
authors.lastname: Lombardi
authors.affiliation: Wolfson Drug Discovery Unit, Centre for Amyloidosis and Acute Phase Proteins, Division of Medicine, University College London, London NW3 2PF, United Kingdom.
authors.forename: Nigel
authors.initials: N
authors.lastname: Rendell
authors.affiliation: Institute of Structural and Molecular Biology, Division of Biosciences, University College London, Gower Street, London WC1E 6BT, United Kingdom.
authors.forename: Mina
authors.initials: M
authors.lastname: Edwards
authors.affiliation: Institute of Structural and Molecular Biology, Division of Biosciences, University College London, Gower Street, London WC1E 6BT, United Kingdom.
authors.forename: Matilda
authors.initials: M
authors.lastname: Katan
authors.affiliation: Proteomics and Molecular Cell Dynamics, Center for Nephrology, School of Life and Medical Sciences, University College London, Royal Free Campus, Rowland Hill Street, London NW3 2PF, United Kingdom.
authors.forename: Jasminka Godovac
authors.initials: JG
authors.lastname: Zimmermann
keywords: Peptides immunoprecipitation; Phosphopeptides enrichment; TMT; Titanium dioxide