Main description:

This volume is a compilation of laboratory protocols and methodology required for the study of molecular chaperones and the cellular stress response. Chapters detail stress response in Hsf1, Hsf2 and Hsf4 knockout mice, mapping HSP interaction networks, the LUminescence-based Mammalian IntERactome (LUMIER), Hsp70 biology, protein folding activity of Hsp90, cytotoxicity of HSP inhibitors, computational approaches for modeling allosteric Hsp90 interactions, HSPs in immunity and vaccine development , and biologies of Hsp70 and Hsp90. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Chaperones: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Contents:

1. Targeted Deletion of Hsf1, 2 and 4 Genes in Mice

Xiongjie Jin, Binnur Eroglu, Demetrius Moskophidis, and Nahid F. Mivechi

2. Role of Heat Shock Factors in Stress-induced Transcription

Murshid, A., Prince T., Ben Lang, and Calderwood, S. K.

3. Monitoring of the Heat Shock Response with a Real-time Luciferase Reporter

Toshiki Kijima, Takanori Eguchi, Len Neckers, and Thomas L. Prince

4. Quantitative Profiling of Chaperone/client Interactions with LUMIER Assay

Mikko Taipale

5. Measurement of chaperone-mediated effects on Polyglutamine Protein Aggregation by the Filter Trap Assay

Maria A.W.H. van Waarde-Verhagen and Harm H. Kampinga

6. Fluorescent-linked Enzyme Chemoproteomic Strategy (FLECS) for Identifying HSP70 Inhibitors

Haystead, T.A.J.

7. A High-throughput Screen for Inhibitors of the Hsp90-Chaperone Machine

Jason Davenport, Lakshmi Galam, and Robert L. Matts

8. Primary Colorectal Cells Culture as a Translation Research Model

Sheah Lin Lee, Nina Claire Dempsey-Hibbert, Dale Vimalachandran, Terence David Wardle, Paul A Sutton, and John H. H. Williams

9. Cell Death and Survival Assays

Alexander E. Kabakov and Vladimir L. Gabai

10. Detecting the Potential Pharmacological Synergy of Drug Combination by Viability Assays In Vitro

Benjamin K. Gibbs and Carole Sourbier

11. Proteomic Profiling of Hsp90 Inhibitors

Sudhakar Voruganti, Jake T. Kline, Maurie J. Balch, Janet Rogers, Robert L. Matts, and Steven D. Hartson

12. Analysis of HspB1 (Hsp27) Oligomerization and Phosphorylation Patterns and its Interaction with Specific Client Polypeptides

Andre-Patrick Arrigo

13. Nucleotide Exchange Factors for Hsp70 Chaperones

Heike Rampelt, Matthias P. Mayer, and Bernd Bukau

14. Determination of Hsp90 Activity through Activation of Glucocorticoid Receptors in Yeast

Michael Reidy

15. Bacterial Hsp90 ATPase Assays

Joel R. Hoskins, Sue Wickner, and Shannon M. Doyle

16. Detecting Post-translational Modifications of Hsp90

Rebecca A. Sager, Mark R. Woodford, Len Neckers, and Mehdi Mollapour

17. Chromatin Immunoprecipitation (ChIP) of Heat Shock Protein 90 (Hsp90)

Aneliya Yoveva and Ritwick Sawarkar

18. A Workflow Guide to RNA-seq Analysis of Chaperone Function and Beyond

Benjamin J. Lang, Kristina M. Holton, Jianlin Gong, and Stuart K. Calderwood

19. Computational Modeling of the Hsp90 Interactions with Cochaperones and Small Molecule Inhibitors

Gennady M. Verkhivker

20. Computational Analysis of the Chaperone Interaction Networks

Shwani Kumar, Kamran Rizzolo, Sandra Zilles, Mohan Babu, and Walid A. Houry

21. Immunohistochemistry of Human Hsp60 in Health and Disease: From Autoimmunity to Cancer

Francesco Cappello, Everly Conway de Macario, Francesca Rappa, Giovanni Zummo, and Alberto J. L. Macario

22. Immunohistochemical and Flow Cytometric Analysis of Intracellular and Membrane-bound Hsp70, as a Putative Biomarker of Glioblastoma Multiforme, using the cmHsp70.1 Monoclonal Antibody

Stefan Stangl, Gemma A. Foulds, Helena Fellinger, Geoffrey J. Pilkington, A. Graham Pockley, and Gabriele Multhoff

23. Detection and Analysis of Extracellular Hsp90 (eHsp90)

Stephanie Cortes, Alexander Baker-Williams, Mehdi Mollapour, and Dimitra Bourboulia

24. Molecular Chaperone Receptors

Ayesha Murshid, Jimmy Theriault, Jianlin Gong, and Stuart K. Calderwood

25. Creation of Recombinant Chaperone Vaccine using Large Heat Shock Protein for Antigen-targeted Cancer Immunotherapy

Chunqing Guo, John R. Subjeck, and Xiang-Yang Wang

26. A Novel Heat Shock Protein 70-based Vaccine Prepared from DC-Tumor Fusion Cells Desheng Weng, Stuart K. Calderwood, and Jianlin Gong

27. Hsp70: A Cancer Target Inside and Outside the Cell Christophe Boudesco, Sebastien Cause, Gaetan Jego, and Carmen Garrido

28. Evidence for Hsp90 Cochaperones in Regulating Hsp90 Function and Promoting Client Protein Folding

Marc B. Cox and Jill L. Johnson

29. Clinical Evaluation and Biomarker Profiling of Hsp90 Inhibitors

Akira Yuno, Min-Jung Lee, Sunmin Lee, Yusuke Tomita, David Rekhtman, Brittni Moore, and Jane B. Trepel