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Phospho-Proteomics

Methods and Protocols

  • Book
  • © 2009

Overview

Editors:
  1. Marjo de Graauw

    1. Division of Toxicology, Leiden/Amsterdam Center for Drug Research (LACDR), Leiden University, Leiden, Netherlands

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  • 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

Part of the book series: Methods in Molecular Biology (MIMB, volume 527)

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Table of contents (23 protocols)

  1. Front Matter

    Pages i-xi
    Download chapter PDF

  2. Protein Phosphorylation and 2D Gel Electrophoresis

    1. A High-Resolution Two Dimensional Gel- and Pro-Q DPS-Based Proteomics Workflow for Phosphoprotein Identification and Quantitative Profiling

      • Ganesh K. Agrawal, Jay J. Thelen
      Pages 3-19

    2. Differential Phosphoprotein Labelling (DIPPL) Using 32P and 33P

      • Aviva M. Tolkovsky, Andreas Wyttenbach
      Pages 21-29

  3. Enrichment of Phosphoproteins and Peptides

    1. Identification of Oxidative Stress-Induced Tyrosine Phosphorylated Proteins by Immunoprecipitation and Mass Spectrometry

      • Ma Carmen Duran, Hong-Lin Chan, John F. Timms
      Pages 33-45

    2. Enrichment and Characterization of Phosphopeptides by Immobilized Metal Affinity Chromatography (IMAC) and Mass Spectrometry

      • Tine E. Thingholm, Ole N. Jensen
      Pages 47-56

    3. The Use of Titanium Dioxide Micro-Columns to Selectively Isolate Phosphopeptides from Proteolytic Digests

      • Tine E. Thingholm, Martin R. Larsen
      Pages 57-66

    4. Enrichment and Separation of Mono- and Multiply Phosphorylated Peptides Using Sequential Elution from IMAC Prior to Mass Spectrometric Analysis

      • Tine E. Thingholm, Ole N. Jensen, Martin R. Larsen
      Pages 67-78

    5. Use of Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) for Phosphotyrosine Protein Identification and Quantitation

      • Guoan Zhang, Thomas A. Neubert
      Pages 79-92

    6. Hydrophilic Interaction Chromatography for Fractionation and Enrichment of the Phosphoproteome

      • Dean E. McNulty, Roland S. Annan
      Pages 93-105

    7. SILAC for Global Phosphoproteomic Analysis

      • Genaro Pimienta, Raghothama Chaerkady, Akhilesh Pandey
      Pages 107-116

    8. Quantitative Phospho-proteomics Based on Soluble Nanopolymers

      • Anton Iliuk, W. Andy Tao
      Pages 117-129

    9. Profiling the Tyrosine Phosphorylation State Using SH2 Domains

      • Kevin Dierck, Kazuya Machida, Bruce J. Mayer, Peter Nollau
      Pages 131-155

  4. Phosphoprotein Analysis by Mass Spectrometry

    1. An Overview of the Qualitative Analysis of Phosphoproteins by Mass Spectrometry

      • Philip R. Gafken
      Pages 159-172

    2. The Analysis of Phosphoproteomes by Selective Labelling and Advanced Mass Spectrometric Techniques

      • Angela Amoresano, Claudia Cirulli, Gianluca Monti, Eric Quemeneur, Gennaro Marino
      Pages 173-190

    3. On-Line Liquid Chromatography Electron Capture Dissociation for the Characterization of Phosphorylation Sites in Proteins

      • Steve M.M. M. Sweet, Helen J. Cooper
      Pages 191-199

    4. Quantification of Protein Phosphorylation by μLC-ICP-MS

      • Ralf Krüger, Nico Zinn, Wolf D. Lehmann
      Pages 201-218

    5. Reverse-Phase Diagonal Chromatography for Phosphoproteome Research

      • Kris Gevaert, Joël Vandekerckhove
      Pages 219-227

    6. Chemical Tagging Strategies for Mass Spectrometry-Based Phospho-proteomics

      • Alexander Leitner, Wolfgang Lindner
      Pages 229-243
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Keywords

About this book

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 Biologyâ„¢ 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.

Editors and Affiliations

  • Division of Toxicology, Leiden/Amsterdam Center for Drug Research (LACDR), Leiden University, Leiden, Netherlands

    Marjo de Graauw

Bibliographic Information

  • Book Title: Phospho-Proteomics

  • Book Subtitle: Methods and Protocols

  • Editors: Marjo de Graauw

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-60327-834-8

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Humana Press 2009

  • Hardcover ISBN: 978-1-60327-833-1Published: 05 March 2009

  • Softcover ISBN: 978-1-61737-917-8Published: 19 November 2010

  • eBook ISBN: 978-1-60327-834-8Published: 25 February 2009

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XII, 332

  • Number of Illustrations: 69 b/w illustrations

  • Topics: Protein Science, Proteomics

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