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Isolation of Heat Shock Protein Complexes

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Molecular Chaperones

Part of the book series: Methods in Molecular Biology ((MIMB,volume 787))

Abstract

Heat shock proteins (Hsp) are molecular chaperones with the capability to interact with a wide range of other proteins and are thus often found coupled with other heat shock and non-heat shock proteins. This can be an advantage to study specific interactions between a chaperone and other proteins and to generate an antitumoral immune response. In this chapter, we present two protocols to isolate Hsp. One involves column chromatography with hydroxyapatite and the other employs immunoprecipitation with antibodies coupled to magnetic beads. In both cases, we specifically want to isolate Hsp coupled with other proteins and use the Hsp complexes as intermediaries to present the coupled peptides/proteins to the immune system, or to explore the associations of a particular Hsp with other proteins.

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References

  1. Dudich IV, Zav’yalov VP, Pfeil W, Gaestel M, Zav’yalova GA, Denesyuk AI, Korpela T (1995) Dimer structure as a minimum cooperative subunit of small heat-shock proteins. Biochemica et Biophysica Acta 1253:163–168

    Article  Google Scholar 

  2. Pinhasi-Kimhi O, Michalovitz D, Ben-Zeev A, Oren M (1986) Specific interaction between the p53 cellular tumour antigen and major heat shock proteins. Nature 320:182–185

    Article  PubMed  CAS  Google Scholar 

  3. Stürzbecher H-W, Addison C, Jenkins JR (1988) Characterization of mutant p53-hsp72/73 protein-protein complexes by transient expression in monkey COS cells. Mol Cell Biol 8:3740–3747

    PubMed  Google Scholar 

  4. Tamura Y, Peng P, Liu K, Daou M, Srivastava PK (1997) Immunotherapy of tumors with autologous tumor-derived heat shock protein preparations. Science 278:117–120

    Article  PubMed  CAS  Google Scholar 

  5. Janetzki S, Palla D, Rosenhauer V, Lochs H, Lewis JJ, Srivastava PK (2000) Immunization of cancer patients with autologous cancer-derived heat shock protein gp96 preparations: A pilot study. Int J Cancer 88:232–238

    Article  PubMed  CAS  Google Scholar 

  6. Eton O, Ross MI, East MJ, Mansfield PF, Papadopoulos N, Ellerhorst JA, Bedikian AY, Lee JE (2010) Autologous tumor-derived heat-shock protein peptide complex-96 (HSPPC-96) in patients with metastatic melanoma. J Transl Med 8:9

    Article  PubMed  Google Scholar 

  7. Wang XY, Kazim L, Repasky EA, Subjeck J (2003) Immunization with tumor-derived ER chaperone grp170 elicits tumor-specific CD8+ T-cell responses and reduces pulmonary metastatic disease. Int J Cancer 105: 226–231

    Article  PubMed  CAS  Google Scholar 

  8. Ciocca DR, Frayssinet P, Cuello-Carrion FD (2007) A pilot study with a therapeutic vaccine based on hydroxyapatite ceramic particles and self-antigens in cancer patients. Cell Stress & Chaperones 12:33–43

    Article  CAS  Google Scholar 

  9. Iyengar R, Rojas F, Rich KA, Birnbaumer L (1991) Membrane receptors: criteria and selected methods of study. In: Hormone Action and Molecular Endocrinology, ed Hughes MR, Schrader WT, O’Malley BW Baylor College of Medicine, Houston, TX, 3–18.

    Google Scholar 

  10. Williams RJP (1989) The functional forms of biominerals. Mann S, Webb J, Williams RJP (Eds), VCH Publisher, Weinheim, 1.

    Google Scholar 

  11. Laquerriere P, Grandjean-Laquerriere A, Jallot E, Balossier G, Frayssinet P, Guenounou M (2003) Importance of hydroxyapatite particles characteristics on cytokines production by human monocytes in vitro. Biomaterials 24: 2739–2747.

    Article  PubMed  CAS  Google Scholar 

  12. Bollag DM, Rozycki MD, Edelstein SJ (1996) Gel electrophoresis under denaturing conditions in Protein Methods (2nd ed) a John Wiley & Sons, Inc, Publication.

    Google Scholar 

  13. Fanelli MA, Cuello-Carrión FD, Dekker J, Schoemaker J, Ciocca DR (1998) Serological detection of heat shock protein hsp27 in normal and breast cancer patients. Cancer Epidemiol Biomarkers Prev 7:791–795.

    PubMed  CAS  Google Scholar 

  14. Bradford, M. M. (1976) A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding. Anal. Biochem. 72:248–254.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

The authors thank Urodelia (Saint Lys, France) for providing the HA.

Competing Interest Statement: The author DRC declare that a patent has been presented on the use of hydroxyapatite for isolation proteins to produce a vaccine. No competing interests are declared by the other authors.

Funding: This work was supported by the grants from the National Research Council (CONICET, PIP 2428), the National Agency for Scientific and Technological Promotion of Argentina (PICT 1047, 2007, Préstamo BID), and the Argentina Foundation for Cancer Research.

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Authors and Affiliations

  1. Laboratory of Oncology, Institute of Experimental Medicine and Biology of Cuyo, Scientific and Technological Center of Mendoza, National Research Council (CONICET), Mendoza, Argentina

    Fernando D. Cuello-Carrión, Mariel A. Fanelli, Niubys Cayado-Gutiérrez, Gisela Castro & Daniel R. Ciocca

  2. Argentina Foundation for Cancer Research, Mendoza, Argentina

    Fernando D. Cuello-Carrión, Mariel A. Fanelli, Niubys Cayado-Gutiérrez, Gisela Castro & Daniel R. Ciocca

Authors
  1. Fernando D. Cuello-Carrión
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  2. Mariel A. Fanelli
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  3. Niubys Cayado-Gutiérrez
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  4. Gisela Castro
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  5. Daniel R. Ciocca
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Corresponding author

Correspondence to Daniel R. Ciocca .

Editor information

Editors and Affiliations

  1. Beth Israel Deaconess Medical Center, Department of Radiation Oncology, Harvard Medical School, Brookline Ave 330, Boston, 02215, Massachusetts, USA

    Stuart K. Calderwood

  2. National Cancer Institute, Urologic Oncology Branch, National Institutes of Health, Rockville Pike 9000, Bethesda, 20814, Maryland, USA

    Thomas L. Prince

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Cuello-Carrión, F.D., Fanelli, M.A., Cayado-Gutiérrez, N., Castro, G., Ciocca, D.R. (2011). Isolation of Heat Shock Protein Complexes. In: Calderwood, S., Prince, T. (eds) Molecular Chaperones. Methods in Molecular Biology, vol 787. Humana Press. https://doi.org/10.1007/978-1-61779-295-3_20

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  • DOI: https://doi.org/10.1007/978-1-61779-295-3_20

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-294-6

  • Online ISBN: 978-1-61779-295-3

  • eBook Packages: Springer Protocols

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