Cell Stress and Chaperones

, Volume 17, Issue 2, pp 145–156 | Cite as

Phage display biopanning identifies the translation initiation and elongation factors (IF1α-3 and eIF-3) as components of Hsp70–peptide complexes in breast tumour cells

  • Christina Siebke
  • Tharappel C. James
  • Robert Cummins
  • Tony O’Grady
  • Elaine Kay
  • Ursula Bond
Original Paper
First Online:
Received:
Revised:
Accepted:

Abstract

The heat shock protein, HSP70, is over-expressed in many tumours and acts at the crossroads of key intracellular processes in its role as a molecular chaperone. HSP70 associates with a vast array of peptides, some of which are antigenic and can mount adaptive immune responses against the tumour from which they are derived. The pool of peptides associated with HSP70 represents a unique barcode of protein metabolism in tumour cells. With a view to identifying unique protein targets that may be developed as tumour biomarkers, we used purified HSP70 and its associated peptide pool (HSP70–peptide complexes, HSP70-PCs) from different human breast tumour cell lines as targets for phage display biopanning. Our results show that HSP70-PCs from each cell line interact with unique sets of peptides within the phage display library. One of the peptides, termed IST, enriched in the biopanning process, was used in a ‘pull-down’ assay to identify the original protein from which the HSP70-associated peptides may have been derived. The eukaryotic translation initiation factor 3 (eIF-3), a member of the elongation factor EF1α family, and the HSP GRP78, were pulled down by the IST peptide. All of these proteins are known to be up-regulated in cancer cells. Immunohistochemical staining of tumour tissue microarrays showed that the peptide co-localised with HSP70 in breast tumour tissue. The data indicate that the reservoir of peptides associated with HSP70 can act as a unique indicator of cellular protein activity and a novel source of potential tumour biomarkers.

Keywords

HSP70-peptide complexes Phage display biopanning Interacting proteins Biomarkers 
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Notes

Acknowledgements

The research was supported by a grant (CFTD/2006/0101) from Enterprise Ireland to UB.

Supplementary material

12192_2011_295_MOESM1_ESM.docx (13 kb)
Supplemental Table 1 (DOCX 12 kb)
12192_2011_295_MOESM2_ESM.docx (11 kb)
Supplemental Table 2 (DOCX 11 kb)

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Copyright information

© Cell Stress Society International 2011

Authors and Affiliations

  • Christina Siebke
    • 1
  • Tharappel C. James
    • 1
  • Robert Cummins
    • 2
  • Tony O’Grady
    • 2
  • Elaine Kay
    • 2
  • Ursula Bond
    • 1
  1. 1.Moyne Institute for Preventive Medicine, School of Genetics and MicrobiologyTrinity College DublinDublin 2Ireland
  2. 2.Royal College of Surgeons in Ireland Education and Research CentreBeaumont HospitalDublin 9Ireland

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