Methods in enzymology. Volume five hundred and twenty seven, Hydrogen peroxide and cell signaling, Part B / edited by Enrique Cadenas and Lester Packer, Pharmacology & Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, CA, USA.

Format:

• Book

Contributor:

• Cadenas, Enrique, editor.
• Packer, Lester, editor.

Series:

• Methods in enzymology ; v. 527.
• Methods in enzymology ; v. 527
• Methods in enzymology ; volume five hundred and twenty seven
• Gale eBooks

Language:

English

Subjects (All):

• Cell interaction.
• Hydrogen peroxide.
• Enzymology.

Physical Description:

1 online resource (lii, 321 pages, 8 unnumbered pages of plates) : illustrations (some color).

Place of Publication:

• Amsterdam ; Boston : Elsevier/AP, 2013.
• San Diego, CA : Academic Press, 2013.

Language Note:

English

Summary:

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

Contents:

• Front Cover; Hydrogen Peroxide and Cell Signaling, Part B; Copyright; Contents; Contributors; Preface; 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; 1. Introduction; 1.1. Measuring H2O2 gradients across the plasma membrane; 1.1.1. Mammalian cell lines; 1.1.2. Yeast cells; 2. Experimental Components and Considerations When Measuring the H2O2 Gradient in S. cerevisiae Cells; Reagents; 2.1. H2O2 calibration curve
• 2.2. Determination of H2O2 consumption by intact and permeabilized yeast cells2.3. Additional points; 3. Experimental Components and Considerations When Measuring the H2O2 Gradient in Mammalian Cell Lines; 3.1. Determination of H2O2 consumption by intact cells; Reagents; 3.2. Determination of H2O2 consumption by disrupted cells; 3.2.1. Preparation of a postnuclear protein extract; 3.2.2. Assay of glutathione peroxidase activity; Reagents; Postnuclear protein extract; Cuvette cell lysate; 3.2.3. Assay of catalase activity; Reagents; In situ; Activity is the slope of the plot.; Cell lysates
• 4. Data Handling/Processing4.1. Yeast cells; 4.2. Cell lines; 5. Summary; Acknowledgments; References; Chapter Two: Evaluating Peroxiredoxin Sensitivity Toward Inactivation by Peroxide Substrates; 1. Introduction; 1.1. Background; 1.2. Theory of the peroxide dependence of sensitivity; 2. Materials; 2.1. Solutions; 2.2. Proteins; 3. Measuring Inactivation Sensitivity by Steady-State NADPH-Linked Assays; 3.1. Evaluation of peroxide sensitivity of E. coli thiol peroxidase with thioredoxin and thioredoxin reductase
• 3.1.1. Demonstration of linear relationship between reaction rate and Prx concentration and establishment of the range of ...3.1.2. Data fitting and analysis; 3.2. Evaluation of substrate specificity for inactivation; 3.2.1. Tpx inactivation by CHP versus H2O2; 3.2.2. Tpx inactivation by CHP in the presence of different reductants (E. coli Trx1 vs. Trx2); 3.2.3. Inactivation of S. typhimurium AhpC by different hydroperoxide substrates; 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 Electrophoresis6. Conclusions/Summary; Acknowledgment; References; Chapter Three: Peroxiredoxins as Preferential Targets in H2O2-Induced Signaling; 1. Introduction; 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

Notes:

• Includes bibliographical references and index.
• "ISSN: 0076-6879."

ISBN:

• 9780124055414
• 0124055419

OCLC:

881096746