It is now understood that the response of mammalian cells to a wide variety of potentially toxic agents may be intimately linked with many human diseases, including rheumatoid arthritis, ischemia, fever, infection, and cancer. In Stress Response: Methods and Protocols, Stephen Keyse has assembled a diverse collection of readily reproducible methods devoted to the study of these varied and powerful responses. Written by leading researchers expert in the techniques they describe, these detailed methods cover the detection and assay of stress-induced damage, the activation of a wide range of signal transduction pathways by cellular stress, stress-induced gene expression, and stress protein function. To ensure experimental success, step-by-step guidance is provided for each method, along with details of reagents, equipment, and other requirements. The methods include both well-established techniques and new technologies at the leading edge of research. Wide ranging and highly practical, Stress Response: Methods and Protocols provides a gold-standard bench manual for today's basic and clinical scientists working to understand how cells and tissues respond during physiological stress and in human disease.

Preface

v

Contributors

xiii

Part I. Detection and Assay of Stress-Induced Damage

Identifying and Counting Protein Modifications Triggered by Nitrosative Stress

3

(12)

Prabodh K. Sehajpal

Harry M. Lander

Determination of Carbonyl Groups in Oxidized Proteins

15

(10)

Rodney L. Levine

Nancy Wehr

Joy A. Williams

Earl R. Stadtman

Emily Shacter

Quantitation of 4-Hydroxynonenal Protein Adducts

25

(10)

Koji Uchida

Earl R. Stadtman

Detection of Oxidative Stress in Lymphocytes Using Dichlorodihydrofluorescein Diacetate

35

(14)

Cecile M. Krejsa

Gary L. Schieven

The Measurement of Protein Degradation in Response to Oxidative Stress

49

(14)

Thomas Reinheckel

Tilman Grune

Kelvin J. A. Davies

Part II. The Activation of Signal Transduction by Cellular Stress

Analysis of the Role of the AMP-Activated Protein Kinase in the Response to Cellular Stress

63

(12)

D. Grahame Hardie

Ian P. Salt

Stephen P. Davies

Detection and Activation of Stress-Responsive Tyrosine Kinases

75

(10)

Gary L. Schieven

Detection of DNA-Dependent Protein Kinase in Extracts from Human and Rodent Cells

85

(14)

Yamini Achari

Susan P. Less-Miller

Expression and Assay of Recombinant ATM

99

(10)

Yael Ziv

Sharon Banin

Dae-Sik Lim

Christine E. Canman

Michael B. Kastan

Yosef Shiloh

Detection and Purification of a Multiprotein Kinase Complex from Mammalian Cells: IKK Signalsome

109

(18)

Frank Mercurio

David B. Young

Anthony M. Manning

Methods to Assay Stress-Activated Protein Kinases

127

(18)

Ana Cuenda

Monitoring the Activation of Stress-Activated Protein Kinases Using GAL4 Fusion Transactivators

145

(16)

Chao-Feng Zheng

Li Xu

Use of Kinase Inhibitors to Dissect Signaling Pathways

161

(16)

Ana Cuenda

Dario R. Alessi

The Development and Use of Phospho-Specific Antibodies to Study Protein Phosphorylation

177

(14)

Jeremy P. Blaydes

Borek Vojtesek

Graham B. Bloomberg

Ted R. Hupp

Peptide Assay of Protein Kinases and Use of Variant Peptides to Determine Recognition Motifs

191

(14)

D. Grahame Hardie

Part III. The Analysis of Stress-Induced Gene Expression

Assaying NF-κB and AP-1 DNA-Binding and Transcriptional Activity

205

(12)

Judith M. Mueller

Heike L. Pahl

Analysis of the Mammalian Heat Shock Response: Inducible Gene Expression and Heat Shock Factor Activity

217

(40)

Anu Mathew

Yanhong Shi

Caroline Jolly

Richard I. Morimoto

Approaches to Define the Involvement of Reactive Oxygen Species and Iron in Ultraviolet-A Inducible Gene Expression

257

(20)

Charareh Pourzand

Olivier Reelfs

Rex M. Tyrrell

The Human Immunodeficiency Virus LTR-Promoter Region as a Reporter of Stress-Induced Gene Expression

277

(20)

Michael W. Bate

Sushma R. Jassal

David W. Brigty

SAGE: The Serial Analysis of Gene Expression

297

(24)

Jill Powell

Analysis of Differential Gene Expression Using the SABRE Enrichment Protocol

321

(26)

Daniel J. Lavery

Phillippe Fonjallaz

Fabienne Fleury-Olela

Ueli Schibler

UVB-Regulated Gene Expression in Human Keratinocytes: Analysis by Differential Display

347

(22)

Harry Frank Abts

Thomas Welss

Kai Breuhahn

Thomas Ruzicka

Part IV. Analysis of Stress Protein Function

Heme Oxygenase Activity: Current Methods and Applications

369

(24)

Stefan W. Ryter

Egil Kvam

Rex M. Tyrrell

Analysis of Molecular Chaperone Activities Using In Vitro and In Vivo Approaches

393

(28)

Brian C. Freeman

Annamieke Michels

Jaewhan Song

Harm H. Kampinga

Richard I. Morimoto

Analysis of Chaperone Properties of Small Hsp's

421

(10)

Monika Ehrnsperger

Matthias Gaestel

Johannes Buchner

Analysis of Small Hsp Phosphorylation

431

(16)

Rainer Benndorf

Katrin Engel

Matthias Gaestel

Analysis of Multisite Phosphorylation of the p53 Tumor-Suppressor Protein by Tryptic Phosphopeptide Mapping

447

(18)

David W. Meek

Diane M. Milne

The Development of Physiological Models to Study Stress Protein Responses

465

(20)

Ted R. Hupp

Index

485

What is included with this book?

The New copy of this book will include any supplemental materials advertised. Please check the title of the book to determine if it should include any access cards, study guides, lab manuals, CDs, etc.

The Used, Rental and eBook copies of this book are not guaranteed to include any supplemental materials. Typically, only the book itself is included. This is true even if the title states it includes any access cards, study guides, lab manuals, CDs, etc.