Phospho-Proteomics

For this volume, top researchers have contributed both established protocols and some of the newest strategies to identify and evaluate protein phosphorylation on Tyr, Ser, and Thr residues, including topics such as 2-dimensional gel electrophoresis.

Protein phosphorylation controls many basic cellular processes, such as cell growth, differentiation, migration, metabolism, and cell death, and its study can provide key insights into the signal transduction pathways that are activated in cells in response to different stimuli, such as growth factor stimulation or exposure to toxicants. In Phospho-Proteomics: Methods and Protocols , expert researchers contribute both well-established protocols and some of the newest strategies for the identification and evaluation of protein phosphorylation on Tyr, Ser, and Thr residues, including topics such as 2-dimensional gel electrophoresis and protein phosphorylation, enrichment of phospho-proteins and peptides, quantitative analysis of phosphorylation by labeling and MS analysis, and antibody and kinase arrays. Written in the highly successful Methods in Molecular Biology™ series format, chapters include brief introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, Phospho-Proteomics: Methods and Protocols is an ideal reference for both new and experienced scientists who wish to gain insight into the current developments in the field and to find inspiration to pursue its future.

Comprehensive phospho-proteomics laboratory manual and research guide for both novice and experienced researchers Provides step-by-step protocols written by leading researchers in the field that are fully adaptable to any specific phospho-proteome analysis task Includes two detailed critical reviews on qualitative and quantitative mass-spectrometry-based phospho-proteomic analysis Covers both basic and state-of-the-art practical procedures as well as bio-informatics approaches to explore the phospho-proteome A compendium of cutting-edge protocols for the identification of phospho-proteins

Inhalt
Protein Phosphorylation and 2D Gel Electrophoresis.- A High-Resolution Two Dimensional Gel- and Pro-Q DPS-Based Proteomics Workflow for Phosphoprotein Identification and Quantitative Profiling.- Differential Phosphoprotein Labelling (DIPPL) Using 32P and 33P.- Enrichment of Phosphoproteins and Peptides.- Identification of Oxidative Stress-Induced Tyrosine Phosphorylated Proteins by Immunoprecipitation and Mass Spectrometry.- Enrichment and Characterization of Phosphopeptides by Immobilized Metal Affinity Chromatography (IMAC) and Mass Spectrometry.- The Use of Titanium Dioxide Micro-Columns to Selectively Isolate Phosphopeptides from Proteolytic Digests.- Enrichment and Separation of Mono- and Multiply Phosphorylated Peptides Using Sequential Elution from IMAC Prior to Mass Spectrometric Analysis.- Use of Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) for Phosphotyrosine Protein Identification and Quantitation.- Hydrophilic Interaction Chromatography for Fractionation and Enrichment of the Phosphoproteome.- SILAC for Global Phosphoproteomic Analysis.- Quantitative Phospho-proteomics Based on Soluble Nanopolymers.- Profiling the Tyrosine Phosphorylation State Using SH2 Domains.- Phosphoprotein Analysis by Mass Spectrometry.- An Overview of the Qualitative Analysis of Phosphoproteins by Mass Spectrometry.- The Analysis of Phosphoproteomes by Selective Labelling and Advanced Mass Spectrometric Techniques.- On-Line Liquid Chromatography Electron Capture Dissociation for the Characterization of Phosphorylation Sites in Proteins.- Quantification of Protein Phosphorylation by ?LC-ICP-MS.- Reverse-Phase Diagonal Chromatography for Phosphoproteome Research.- Chemical Tagging Strategies for Mass Spectrometry-Based Phospho-proteomics.- Arrays to StudyProtein-Phosphorylation.- Antibody Array Platform to Monitor Protein Tyrosine Phosphorylation in Mammalian Cells.- Protein Tyrosine Kinase Characterization Based on Fully Automated Synthesis of (Phospho) Peptide Arrays in Microplates.- Kinome Profiling Using Peptide Arrays in Eukaryotic Cells.- Bioinformatics.- ProMoST: A Tool for Calculating the pI and Molecular Mass of Phosphorylated and Modified Proteins on Two-Dimensional Gels.- Kinase-Specific Prediction of Protein Phosphorylation Sites.- Reconstructing Regulatory Kinase Pathways from Phosphopeptide Data: A Bioinformatics Approach.