Redox-Mediated Signal Transduction

by Hancock, John T.

• ISBN13: 9781588298423
• ISBN10: 1588298426
• Edition: 1st
• Format: Hardcover
• Copyright: 2008-10-04
• Publisher: Humana Pr Inc

Summary

Once believed to be involved mainly with energetics, including the production of ATP, knowledge of the role of redox in the control of cellular activity has been expanded over recent years. In Redox-Mediated Signal Transduction: Methods and Protocols, experienced researchers with backgrounds in both the plant and animal sciences contribute timely methods and techniques that can be used to study this important aspect of biology. Beginning with an overview and methods for measuring compounds that affect redox and the redox state of cells, the book continues with reviews of the use of GFP and its derivatives, methods to study the impact of changing redox on proteins, and methods to study the exact molecular changes that may underlie the mechanisms of action of altering redox, among other subjects. As a volume in the highly successful Methods in Molecular Biology series, chapters include step-by-step, readily reproducible protocols, lists of the necessary materials and reagents, and tips on troubleshooting and avoiding known pitfalls.

Table of Contents

Preface	p. v
Contributors	p. ix
The Role of Redox in Signal Transduction	p. 1
The Measurement of Nitric Oxide and its Metabolites in Biological Samples by Ozone-based Chemiluminescence	p. 11
Detection and Measurement of Reactive Oxygen Intermediates in Mitochondria and Cells	p. 29
Redox-sensitive Green Fluorescent Protein: Probes for Dynamic Intracellular Redox Responses. A Review	p. 51
Measuring Redox Changes in Vivo in Leaves: Prospects and Technical Challenges	p. 67
Imaging of Intracellular Hydrogen Peroxide Production with Hyper Upon Stimulation of Hela Cells with Egf	p. 79
Tools to Investigate ROS Sensitive Signalling Proteins	p. 87
Methods for Preparing Crystals of Reversibly Oxidized Proteins: Crystallization of Protein Tyrosine Phosphatase 1b as an Example	p. 101
Methods for the Study of Redox-mediated Changes in p53 Structure and Function	p. 117
Redox Regulation and Trapping Sulphenic Acid in the Peroxide Sensitive Human Mitochondrial Branched Chain Aminotransferase	p. 139
Detection of Carbonylated Proteins in 2-D SDS Page Separations	p. 153
Analysis of Global and Specific Changes in the Disulfide Proteome Using Redox 2D-PAGE	p. 165
Protein-thiol Oxidation, from Single Proteins to Proteome-wide Analyses	p. 181
Analysis of Redox Relationships in the Plant Cell Cycle: Determinations of Ascorbate, Glutathione and Poly (ADPribose) Polymerase (PARP) in Plant Cell Cultures	p. 199
Generation and Detection of S-Nitrosothiols	p. 217
Index	p. 231
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