Contents

Preface v

Contributors ix

1 The Role of Redox in Signal Transduction 1

John T. Hancock

2 The Measurement of Nitric Oxide and its Metabolites in Biological Samples by Ozone-based Chemiluminescence 11

Andrew G. Pinder, Stephen C. Rogers, Afshin Khalatbari, Thomas E. Ingram, and Philip E. James.

3 Detection and Measurement of Reactive Oxygen Intermediates in Mitochondria and Cells 29

Matthew Whiteman, Yuktee Dogra, Paul G. Winyard, and Jeffrey S. Armstrong

4 Redox-sensitive Green Fluorescent Protein: Probes for Dynamic Intracellular Redox Responses. A Review 51

Mark B. Cannon and S. James Remington

5 Measuring Redox Changes in Vivo in Leaves: Prospects and Technical Challenges 67

Philip M. Mullineaux and Tracy Lawson

6 Imaging of Intracellular Hydrogen Peroxide Production with Hyper Upon Stimulation of Hela Cells with Egf 79

Kseniya N. Markvicheva, Ekaterina A. Bogdanova,

Dmitry B. Staroverov, Sergei Lukyanov, and Vsevolod V. Belousov

7 Tools to Investigate ROS Sensitive Signalling Proteins 87

Radhika Desikan, Steven J. Neill, Jenna Slinn, and John T. Hancock

8 Methods for Preparing Crystals of Reversibly Oxidized Proteins:

Crystallization of Protein Tyrosine Phosphatase lb as an Example 101

Annette Salmeen and David Barford

9 Methods for the Study of Redox-mediated Changes in p53 Structure and Function 117

Kristine Mann

10 Redox Regulation and Trapping Sulphenic Acid in the Peroxide Sensitive Human Mitochondrial

Branched Chain Aminotransferase 139

Susan M Hutson, Leslie B. Poole, Steven Coles, and Myra E. Conway

11 Detection of Carbonylated Proteins in 2-D SDS

Page Separations 153

Rukhsana Sultana, Shelley F. Newman, Quanzhen Huang, and D. Allan Butterfield

12 Analysis of Global and Specific Changes in the Disulfide

Proteome Using Redox 2D-PAGE 165

Robert C. Cumming 13 Protein-thiol Oxidation, from Single Proteins to Proteome-wide Analyses 181

Natacha Le Moan, Frederique Tacnet, and Michel B. Toledano

14 Analysis of Redox Relationships in the Plant Cell Cycle:

Determinations of Ascorbate, Glutathione and Poly (ADPribose)

Polymerase (PARP) in Plant Cell Cultures 199

Christine H. Foyer, Till K. Pellny, Vittoria Locato, and Laura De Gara

15 Generation and Detection of S-Nitrosothiols 217

Christian Lindermayr, Simone Sell, and Jörg Durner

Index 231

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