Analytical strategies in mass spectrometry-based phosphoproteomics

Heidi Rosenqvist, Juanying Ye, Ole N Jensen

Research output: Contribution to journalJournal article

Abstract

Phosphoproteomics, the systematic study of protein phosphorylation events and cell signaling networks in cells and tissues, is a rapidly evolving branch of functional proteomics. Current phosphoproteomics research provides a large toolbox of strategies and protocols that may assist researchers to reveal key regulatory events and phosphorylation-mediated processes in the cell and in whole organisms. We present an overview of sensitive and robust analytical methods for phosphopeptide analysis, including calcium phosphate precipitation and affinity enrichment methods such as IMAC and TiO(2). We then discuss various tandem mass spectrometry approaches for phosphopeptide sequencing and quantification, and we consider aspects of phosphoproteome data analysis and interpretation. Efficient integration of these stages of phosphoproteome analysis is highly important to ensure a successful outcome of large-scale experiments for studies of phosphorylation-mediated protein regulation.
Original languageEnglish
JournalMethods in Molecular Biology
Volume753
Pages (from-to)183-213
Number of pages31
ISSN1064-3745
DOIs
Publication statusPublished - 1. Jan 2011

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