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METHODS IN ENZYMOLOGY
Methods in Enzymology
Section I: H2O2 Metabolism: Determination of a Cellular Steady-State
Chapter One. The Cellular Steady-State of H2O2: Latency Concepts and Gradients
2 Experimental Components and Considerations When Measuring the H2O2 Gradient in S. cerevisiae Cells
3 Experimental Components and Considerations When Measuring the H2O2 Gradient in Mammalian Cell Lines
4 Data Handling/Processing
Chapter Two. Evaluating Peroxiredoxin Sensitivity Toward Inactivation by Peroxide Substrates
3 Measuring Inactivation Sensitivity by Steady-State NADPH-Linked Assays
4 Measuring Inactivation Sensitivity by Multiturnover Cycling with ROOH and DTT Followed by Mass Spectrometry Analysis
5 Measuring Inactivation Sensitivity of Prx1/AhpC Prxs Under Single Turnover Conditions Followed by Gel Electrophoresis
Chapter Three. Peroxiredoxins as Preferential Targets in H2O2-Induced Signaling
2 Reaction of H2O2 with Cellular Thiols
3 H2O2 Diffusion Versus Reaction with Cellular Thiols
4 Sulfenic Acids as Signal Transduction Intermediates
5 Prxs as Preferential Targets for H2O2
6 Prxs as Primary H2O2 Sensors and Transducers
7 Prx–Protein Interactions are Needed to Transmit the Signal
8 Posttranslational Regulation of Prxs
Chapter Four. Selenium in the Redox Regulation of the Nrf2 and the Wnt Pathway
1 Some Historical Background for Introduction
2 Selenium Status and Selenoprotein Synthesis
3 Selenium Status and the Keap1/Nrf2 System
4 Selenium and the Wnt Pathway
5 Common Players and Events in Nrf2 and Wnt Signaling
6 How Does Selenium Come into Play?
Chapter Five. Selenoprotein W as Biomarker for the Efficacy of Selenium Compounds to Act as Source for Selenoprotein Biosynthesis
Chapter Six. Peroxiredoxins and Sulfiredoxin at the Crossroads of the NO and H2O2 Signaling Pathways
2 The Effect of NO on the Level of 2-Cys-Prx Overoxidation
3 Detection of Srx
Chapter Seven. Glutathione and γ-Glutamylcysteine in Hydrogen Peroxide Detoxification
3 Previous Considerations
4 Procedure with Purified Enzymes and Substrates
5 Analysis in Biological Samples
6 Further Applications: H2O2 Produced by NOS
Chapter Eight. Peroxiredoxin-6 and NADPH Oxidase Activity
2 Experimental Components and Considerations
3 Peroxiredoxin Activity of Prdx6
4 Effect of Prdx6 on NADPH Oxidase Activity
Chapter Nine. Study of the Signaling Function of Sulfiredoxin and Peroxiredoxin III in Isolated Adrenal Gland: Unsuitability of Clonal and Primary Adrenocortical Cells
2 Hyperoxidation of PrxIII by H2O2 Generated During Corticosterone Synthesis
3 Induction of Srx by ACTH
4 Unsuitability of Clonal and Primary Adrenocortical Cells for Studies of the Srx–PrxIII Regulatory Pathway
5 Adrenal Gland Organ Culture as an In Vitro Model for the Srx–PrxIII Regulatory Pathway
6 Concluding Remarks
Section II: H2O2 in the Regulation of Cellular Processes in Plants
Chapter Ten. The Use of HyPer to Examine Spatial and Temporal Changes in H2O2 in High Light-Exposed Plants
2 Experimental Procedures
3 Pilot Experiments Using HL Stress
Chapter Eleven. A Simple and Powerful Approach for Isolation of Arabidopsis Mutants with Increased Tolerance to H2O2-Induced Cell Death
2 Generation and Isolation of Mutants More Tolerant to H2O2-Induced Oxidative Stress
3 Identification of Mutations in the Genome
4 Analysis of the Mutants with Enhanced Tolerance to H2O2-Induced Oxidative Stress
Chapter Twelve. Analysis of Environmental Stress in Plants with the Aid of Marker Genes for H2O2 Responses
2 Experimental Materials and Procedures
3 Example of Analysis
Chapter Thirteen. The Role of Plant Bax Inhibitor-1 in Suppressing H2O2-Induced Cell Death
2 Morphological Changes of Mitochondria Under ROS Stress
3 Assay for Inhibitory Effect of BI-1 on ROS Stress-Induced Cell Death Using Heterologous Expression System in Suspension Cultured Cells
Chapter Fourteen. Comparative Analysis of Cyanobacterial and Plant Peroxiredoxins and Their Electron Donors: Peroxidase Activity and Susceptibility to Overoxidation
2 Expression and Purification of Recombinant Prxs and Thioredoxins
3 Prx Activity Assays In Vitro
4 Peroxide Decomposition in Cyanobacteria In Vivo
5 Overoxidation of Plant and Cyanobacterial 2-Cys Prx
6 Concluding Remarks
Chapter Fifteen. Using Hyper as a Molecular Probe to Visualize Hydrogen Peroxide in Living Plant Cells: A Method with Virtually Unlimited Potential in Plant Biology
2 NADPH Oxidase in Plant Cells
3 Plant Cells Respond to External and Internal Stimuli
4 Visualizing Hydrogen Peroxide in Living Plant Cells
5 Hyper as a New Genetically Encoded Probe
6 Vector Description and Plant Transformation
7 Preparation and Sterilization of Modified Petri Dishes for Growing Arabidopsis Plants for Microscopy Analysis
8 Seeds Sterilization and Stratification
9 Growth Conditions
10 Image Acquisition and Processing
This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This is the second of three volumes on hydrogen peroxide and cell signaling, and includes chapters on such topics as the cellular steady-state of H2O2, evaluating peroxiredoxin sensitivity towards inactivation by peroxide substrates, and peroxiredoxins as preferential targets in H2O2-induced signaling.
- Continues the legacy of this premier serial with quality chapters authored by leaders in the field
- Covers hydrogen peroxide and cell signaling
- Contains chapters on such topics as the cellular steady-state of H2O2, evaluating peroxiredoxin sensitivity towards inactivation by peroxide substrates, and peroxiredoxins as preferential targets in H2O2-induced signaling
Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists.
- No. of pages:
- © Academic Press 2013
- 15th July 2013
- Academic Press
- Hardcover ISBN:
- eBook ISBN:
Lester Packer received a PhD in Microbiology and Biochemistry in 1956 from Yale University. In 1961, he joined the University of California at Berkeley serving as Professor of Cell and Molecular Biology until 2000, and then was appointed Adjunct Professor, Pharmacology and Pharmaceutical Sciences, School of Pharmacy at the University of Southern California.
Dr Packer received numerous distinctions including three honorary doctoral degrees, several distinguished Professor appointments. He was awarded Chevalier de l’Ordre National du Merite (Knight of the French National Order of Merit) and later promoted to the rank of Officier. He served as President of the Society for Free Radical Research International (SFRRI), founder and Honorary President of the Oxygen Club of California.
He has edited numerous books and published research; some of the most cited articles have become classics in the field of free radical biology:
Dr Packer is a member of many professional societies and editorial boards. His research elucidated - the Antioxidant Network concept. Exogenous lipoic acid was discovered to be one of the most potent natural antioxidants and placed as the ultimate reductant or in the pecking order of the “Antioxidant Network” regenerating vitamins C and E and stimulating glutathione synthesis, thereby improving the overall cellular antioxidant defense. The Antioxidant Network is a concept addressing the cell’s redox status. He established a world-wide network of research programs by supporting and co-organizing conferences on free radical research and redox biology in Asia, Europe, and America.
Department of Molecular Pharmacology and Toxicology, School of Pharmaceutical Sciences, University of Southern California, USA
ENRIQUE CADENAS, MD, PhD, received his PhD in biochemistry from the University of Buenos Aires, School of Medicine. He is professor of pharmacology and pharmaceutical sciences at the University of Southern California School of Pharmacy and of biochemistry and molecular biology at the University of Southern California Keck School of Medicine, and doctor honoris causa (medicine) at the University of Linköping, Sweden. Cadenas was president of the Society for Free Radical Research International (SFRRI) and is fellow of the Society for Free Radical Biology & Medicine. He served the scientific community by participating on NIH study sections (2002-2006; chair 2006-2008). His research interests include energy and redox metabolism in brain aging and the coordinated inflammatory-metabolic responses in brain and neurodegenerative diseases.
Pharmacology & Pharmaceutical Sciences, School of Pharmacy, University of Southern California, USA
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