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Protein Sensors of Reactive Oxygen Species, Part A: Selenoproteins and Thioredoxin
1st Edition, Volume 347 - March 1, 2002
Editors: Lester Packer, Helmut Sies
Language: English
Hardback ISBN:9780121822484
9 7 8 - 0 - 1 2 - 1 8 2 2 4 8 - 4
eBook ISBN:9780080496955
9 7 8 - 0 - 0 8 - 0 4 9 6 9 5 - 5
This volume of Methods in Enzymology is concerned with the rapidly developing field of selenoprotein synthesis and its related molecular genetics. Progressive information on the to…Read more
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This volume of Methods in Enzymology is concerned with the rapidly developing field of selenoprotein synthesis and its related molecular genetics. Progressive information on the topics of proteins as redox sensors, selenoproteins, and the thioredoxin system is studied using methods such as bioinformatics, DNA chip technology, cell biology, molecular genetics, and enzymology. The information on novel selenoproteins identified from genomic sequence data, as well as current knowledge on glutathione peroxidases, selenoprotein P, iodothyronine deiodinases, and thioredoxin reductases, is presented in a method-based approach.
Biochemists, pharmacologists, physiologists, cell biologists, molecular biologists, and biomedical researchers.
Contributors to volume 347
Preface
Methods in enzymology
Section I: Selenoproteins
[1]: Selenoprotein Biosynthesis: Purification and Assay of Components Involved in Selenocysteine Biosynthesis and Insertion in Escherichia coli
Introduction
Purification of Selenocysteine-Inserting tRNASec
Purification of Selenophosphate Synthetase
Purification of Selenocysteine Synthase
Preparation of Elongation Factor SelB
Serylation of tRNASec
Conversion of Seryl-tRNASec to Selenocysteyl-tRNASec
Preparation of Selenocysteine Insertion Sequence Elements
Analysis of Interaction of SelB with Its RNA Ligands
[2]: Selenocysteine Insertion Sequence Element Characterization and Selenoprotein Expression
Abstract
Criteria for Identifying Functional Selenocysteine Insertion Sequence Elements in Databases
Methods for Studying Selenocysteine Insertion Sequence Function and Selenoprotein Synthesis
Selenocysteine Insertion Sequence Activity Assays
Optimizing Expression in Transiently Transfected Cells
Incorporation of 75Se into Selenoproteins in Transiently Transfected Cells
Acknowledgment
[3]: Transfer RNAs That Insert Selenocysteine
Introduction
Primary Structures
Selenocysteine-Inserting tRNA Gene, Gene Expression, and Gene Product
Selenocysteine-Inserting tRNA: Carrier Molecule for Biosynthesis of Its Amino Acid
Selenocysteine-Inserting tRNA: Donor of Selenocysteine to Protein
Functional Consequences of Over- and Underexpression of Mammalian Selenocysteine-Inserting tRNA and Expression of an i 6 A-Deficient Selenocysteine-Inserting tRNA
Purification of Selenocysteine-Inserting tRNA
Preparation of [75Se]Selenocysteyl-tRNA
[4]: Purification and Analysis of Selenocysteine Insertion Sequence-Binding Protein 2
Introduction
Purification of Native Selenocysteine Insertion Sequence-Binding Protein 2
Purification of Recombinant Selenocysteine Insertion Sequence-Binding Protein 2
RNA-Binding Assays for Selenocysteine Insertion Sequence-Binding Protein 2
In Vitro Translation of Phospholipid Hydroperoxide Glutathione Peroxidase
Ribosome-Binding Assay
Summary and Perspectives
Acknowledgments
[5]: Nonsense-Mediated Decay: Assaying for Effects on Selenoprotein mRNAs
Introduction
Rationale for Analyzing Transiently Expressed, in Vitro-Modified Glutathione Peroxidase 1 Alleles in Cultured Cells
Methods Used to Determine the Mechanism by Which Selenium Concentration Affects Glutathione Peroxidase 1 Gene Expression
Summary
Acknowledgment
[6]: Novel Selenoproteins Identified from Genomic Sequence Data
Introduction
Computational Detection of Selenocysteine Insertion Sequence Motifs
Functional Screen of Selected Selenocysteine Insertion Sequence Hits
Obtaining and Analyzing Selenoprotein cDNA Sequences
Further Characterization of Selenoproteins
Conclusion
Acknowledgments
[7]: Semisynthesis of Proteins Containing Selenocysteine
Introduction
Strategies for Synthesis of Proteins Containing Selenocysteine
Preparation of Selenocysteine for Peptide Synthesis
Solid-Phase Synthesis of Peptide Containing Selenocysteine
Semisynthesis of a Protein Containing Selenocysteine
Characterization of Proteins Containing Selenocysteine
Use of Selenocysteine as Cleavage Reagent
Summary
Acknowledgments
[8]: Mammalian Selenoprotein Gene Signature: Identification and Functional Analysis of Selenoprotein Genes Using Bioinformatics Methods
Introduction
Eukaryotic Selenoproteins
Selenocysteine/Cysteine Pair in Homologous Sequences
Mammalian Selenocysteine Insertion Sequence Element
Identification of Selenocysteine Insertion Sequence Elements
Mammalian Selenoprotein Gene Signature
Applicability of Mammalian Selenoprotein Gene Signature Criteria
Other Approaches for Identification of Selenoprotein Genes
Strategy to Search for Selenoproteins in Nucleotide Sequence Databases
Strategy to Test Whether a Newly Isolated Gene Encodes a Selenoprotein
Functional Analyses of New Selenoproteins
Concluding Remarks
Acknowledgment
[9]: Estimation of Individual Types of Glutathione Peroxidases
Introduction
Determination by Activity
Determination by RNA Analysis
Determination by Immunochemical Methods
Immunochemical Detection
Conclusions
Acknowledgments
[10]: High-Throughput 96-Well Microplate Assays for Determining Specific Activities of Glutathione Peroxidase and Thioredoxin Reductase
Introduction
Methods
[11]: Selenoprotein P
Introduction
Structure
Properties
Proposed Functions
Acknowledgments
[12]: Iodothyronine Deiodinases
Introduction
Historical Reminiscence
Type I 5′-Deiodinase
Type II 5′-Deiodinase Activity
Type III Iodothyronine Deiodinase
Methods to Determine Deiodinase Activity
Acknowledgments
[13]: Expression and Regulation of Thioredoxin Reductases and Other Selenoproteins in Bone
Bone Physiology
Expression of Selenoproteins in Cells of Bone Microenvironment
Expression and Regulation of Glutathione Peroxidases in Osteoblasts
Expression and Regulation of Thioredoxin Reductases in Osteoblasts
Expression of Glutathione Peroxidases and Thioredoxin Reductase α in Monocyte-Derived Cells
Discussion
Summary
[14]: Selenoprotein W
Abstract
Procedures
[15]: Genetic and Functional Analysis of Mammalian Sep15 Selenoprotein
Introduction
Materials and Methods
Conclusions
Acknowledgment
[16]: Selenocysteine Lyase from Mouse Liver
Introduction
Cloning of cDNA for Mouse Selenocysteine Lyase
Expression of Selenocysteine Lyase in Escherichia coli
Assay Method
Purification Procedure
Properties
Tissue Distribution and Intracellular Localization
[17]: Selenocysteine Methyltransferase
Introduction
Enzyme Assays
Semiquantitative Assay
Quantitative Assay
Crude Extract and Ammonium Sulfate Precipitation
Column Chromatography
Storage and Stability
Properties
Relation to Homocysteine Methyltransferases
[18]: Phospholipid–Hydroperoxide Glutathione Peroxidase in Sperm
Introduction
Measurement of Phospholipid-Hydroperoxide Glutathione Peroxidase in Human Spermatozoa
Discussion
Acknowledgments
[19]: In Vivo Antioxidant Role of Glutathione Peroxidase: Evidence from Knockout Mice
Introduction
Development and Characterization of GPX1(+) and GPX1(−/−) Mouse Models
Protection by Glutathione Peroxidase against Acute, Lethal Oxidative Stress
Protection by Glutathione Peroxidase against Moderate and Metabolic Oxidative Stress
Unanswered Questions
Acknowledgment
[20]: Recombinant Expression of Mammalian Selenocysteine-Containing Thioredoxin Reductase and Other Selenoproteins in Escherichia coli
Introduction
tRNASec Defining the Selenoprotein World
Species Barriers in Heterologous Selenoprotein Synthesis
Introduction of Selenocysteine Insertion Sequence Element Compatible with Bacterial Selenoprotein Synthesis Machinery as Method for Recombinant Selenoprotein Production
Critical Factors for Recombinant Selenoprotein Production in Escherichia coli
Conclusions
[21]: Mammalian Thioredoxln Reductases as Hydroperoxide Reductases
Introduction
Purification of Mammalian Thioredoxin Reductase from Wet Tissue
Assay of Mammalian Thioredoxin Reductase Activity
Assay of Hydroperoxide Reductase Activity
Discussion
Comments
Acknowledgments
[22]: Tryparedoxin and Tryparedoxin Peroxidase
Introduction
Tryparedoxins
Tryparedoxin Peroxidases
Sources of Purified Tryparedoxin and Tryparedoxin Peroxidase
Conclusions
Acknowledgments
[23]: Trypanothione and Tryparedoxin in Ribonucleotide Reduction
Introduction
Methods
Conclusions
Acknowledgment
[24]: Selenium- and Vitamin E-Dependent Gene Expression in Rats: Analysis of Differentially Expressed mRNAs
Introduction
Production of Selenium and Vitamin E Deficiency in Rats
Analysis of Differentially Expressed mRNAs by Atlas cDNA Expression Arrays
Typical Results
Section II: Thioredoxin
[25]: Overview
Introduction
Thioredoxin and Related Molecules
Thioredoxin-Binding Proteins
Redox Regulation of Transcriptional Factors by Thioredoxin
Cytoprotective Action of Thioredoxin
Environmental Stressors and Redox Signal
Concluding Remarks and Perspectives
Acknowledgments
[26]: Thioredoxin and Glutaredoxin Isoforms
Introduction
Thioredoxins
Escherichia coli Thioredoxins
Saccharomyces cerevisiae Thioredoxins
Human Thioredoxins
Structural Similarities
Assays for Thioredoxin Activity
Glutaredoxins
Catalytic Mechanism of Glutaredoxin/Thioredoxin
Escherichia coli Glutaredoxins
Saccharomyces cerevisiae Glutaredoxins
Human Glutaredoxin
Classification of Glutaredoxins
Assays for Glutaredoxin Activity
Acknowledgments
[27]: Mammalian Thioredoxin Reductases
Introduction
Labeling Proteins with Selenium-75
Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis Analysis and Autoradiography
Identification of Selenocysteine Residues
Precautions for Purifying Selenocysteine-Containing Proteins
Separation of Thioredoxin Reductase 1 from Thioredoxin Reductase 2 in Rat Liver Homogenates
Selective Alkylation of Selenocysteine Residue 498 of Thioredoxin Reductase 1
Catalytic Role of Selenocysteine Residue
[28]: Mitochondrial Thioredoxin Reductase and Thiol Status
Introduction
Determination of Total Mitochondrial Thiols
Determination of Total Protein Thiols, Membrane Thiols, and Acid-Soluble Thiols
Purification of Mitochondrial Thioredoxin Reductase
Preparation and Processing of Mitochondria
DEAE-Sephacel Chromatography
Assays
Acknowledgment
[29]: Protein Electrophoretic Mobility Shift Assay to Monitor Redox State of Thioredoxin in Cells
Introduction
Methods
Results
Acknowledgments
[30]: Recycling of Vitamin C by Mammalian Thioredoxin Reductase
Introduction
Methods
Conclusion
[31]: Thioredoxin Cytokine Action
Introduction
Truncated Form of Thioredoxin
Thioredoxin as Costimulatory Molecule of Cytokine Action
Chemokine-Like Activity of Thioredoxin
Circulating Thioredoxin Levels in Human Plasma
Sandwich Enzyme-Linked Immunosorbent Assay for Human Thioredoxin
Concluding Remarks
Acknowledgment
[32]: Identification of Thioredoxin-Linked Proteins by Fluorescence Labeling Combined with Isoelectric Focusing/Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis
Introduction
Sources of Materials and Chemicals
Methods
Applications
[33]: Thioredoxin and Mechanism of Inflammatory Response
Introduction
NF-κB Immunostaining with Synovial Fibroblast Cultures
Electrophoretic Mobility Shift Assay
[34]: Redox State of Cytoplasmic Thioredoxin
Introduction
Determination of Redox State in Vivo
Thioredoxin as Disulfide Bond Reductase
Thioredoxin as Thiol Oxidant
Acknowledgments
[35]: Thioredoxin, Thioredoxin Reductase, and Thioredoxin Peroxidase of Malaria Parasite Plasmodium falciparum
Introduction
Expression and Purification of Recombinant Thioredoxin Reductase, Thioredoxin 1, and Thioredoxin Peroxidase 1
Assays for Reactions and Compounds of Thioredoxin System
Concluding Remarks
Acknowledgments
[36]: Human Placenta Thioredoxin Reductase: Preparation and Inhibitor Studies
Introduction
Assay Procedures
[37]: Classification of Plant Thioredoxins by Sequence Similarity and Intron Position
Arabidopsis Genes Encoding Thioredoxins or Proteins with Thioredoxin Domains
Usability of Arabidopsis Data for Predicting Type of Plant Sequence
[38]: Ferredoxin-Dependent Thioredoxin Reductase: A Unique Iron–Sulfur Protein
Introduction
Ferredoxin/Thioredoxin Reductase
Spinach Thioredoxin f
Acknowledgment
[39]: Plant Thioredoxin Gene Expression: Control by Light, Circadian Clock, and Heavy Metals
Introduction
Experimental Procedures
[40]: Thioredoxin Genes in Lens: Regulation by Oxidative Stress
Introduction
Experimental Procedures
mRNAs Encoding Thioredoxin Genes in Lens
Western Blot Analysis of Thioredoxin
Thioredoxin Reductase Activity in Human Lens
Concluding Remarks
Acknowledgments
[41]: Thioredoxin Overexpression in Transgenic Mice
Introduction
Thioredoxin Knockout Mice
Characteristics of Thioredoxin-Transgenic Mice
Concluding Remarks
Acknowledgment
[42]: Multiplex Reverse Transcription-Polymerase Chain Reaction for Determining Transcriptional Regulation of Thioredoxin and Glutaredoxin Pathways
Introduction
RNA
cDNA
Primers
Multiplex Polymerase Chain Reaction
Optimization of Multiplex Reverse Transcription-Polymerase Chain Reaction
Application
Final Remarks
Acknowledgments
[43]: Redox Regulation of Cell Signaling by Thioredoxin Reductases
Introduction
Isolation of Mammalian Thioredoxin Reductase Isozymes
Analysis of Thioredoxin Reductase 1 Redox State, Using 5-Iodoacetamidofluorescein
Analysis of Changes in Redox State of Isolated Mouse Liver Thioredoxin Reductase 1 in Vitro
Analysis of Thioredoxin Reductase 1 Redox State in Cell Culture System
Specific Alkylation of Selenocysteine in Thioredoxin Reductase 1 by 5-Iodoacetamidofluorescein
Effect of Stimulation of A431 Cells with Epidermal Growth Factor or H2O2 on Expression of Thioredoxin Reductase 1
Concluding Remarks
Author index
Subject index
No. of pages: 511
Language: English
Edition: 1
Volume: 347
Published: March 1, 2002
Imprint: Academic Press
Hardback ISBN: 9780121822484
eBook ISBN: 9780080496955
LP
Lester Packer
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.
Affiliations and expertise
Department of Molecular Pharmacology and Toxicology, School of Pharmaceutical Sciences, University of Southern California, USA
HS
Helmut Sies
Helmut Sies, MD, PhD (hon), studied medicine at the universities of Tübingen, Munich, and Paris. He was the professor and chair of the Institute for Biochemistry and Molecular Biology I at Heinrich-Heine-University Düsseldorf, Germany, where he is now professor emeritus. He is a member of the German National Academy of Sciences Leopoldina and was the president of the North Rhine-Westphalian Academy of Sciences and Arts. He was named ‘Redox Pioneer’; was the president of the Society for Free Radical Research International (SFRRI). Helmut Sies introduced the concept of “Oxidative Stress” in 1985, and was the first to reveal hydrogen peroxide as a normal constituent of aerobic cell metabolism. His research interests comprise redox biology, oxidants, antioxidants, micronutrients.
Affiliations and expertise
Heinrich-Heine-University Düsseldorf, Germany
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