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A high-affinity conformation of Hsp90 confers tumour selectivity on Hsp90 inhibitors

Nature volume 425, pages 407–410 (2003)Cite this article

Abstract

Heat shock protein 90 (Hsp90) is a molecular chaperone that plays a key role in the conformational maturation of oncogenic signalling proteins, including HER-2/ErbB2, Akt, Raf-1, Bcr-Abl and mutated p531,2,3,4,5,6,7. Hsp90 inhibitors bind to Hsp90, and induce the proteasomal degradation of Hsp90 client proteins6,8,9,10,11. Although Hsp90 is highly expressed in most cells, Hsp90 inhibitors selectively kill cancer cells compared to normal cells, and the Hsp90 inhibitor 17-allylaminogeldanamycin (17-AAG) is currently in phase I clinical trials12,13. However, the molecular basis of the tumour selectivity of Hsp90 inhibitors is unknown. Here we report that Hsp90 derived from tumour cells has a 100-fold higher binding affinity for 17-AAG than does Hsp90 from normal cells. Tumour Hsp90 is present entirely in multi-chaperone complexes with high ATPase activity, whereas Hsp90 from normal tissues is in a latent, uncomplexed state. In vitro reconstitution of chaperone complexes with Hsp90 resulted in increased binding affinity to 17-AAG, and increased ATPase activity. These results suggest that tumour cells contain Hsp90 complexes in an activated, high-affinity conformation that facilitates malignant progression, and that may represent a unique target for cancer therapeutics.

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Figure 1: 17-AAG has a higher binding affinity to Hsp90 from tumour cells than normal cells or purified Hsp90 protein.
Figure 2: Hsp90 from tumour cells is present in chaperone complexes with high ATPase activity.
Figure 3: In vitro reconstitution of purified Hsp90 with co-chaperones increased binding affinity to 17-AAG and the ATPase activity.
Figure 4: Hsp90 from clinical tumour samples is in a high-affinity complex with increased ATPase activity.

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References

  1. Schulte, T. W. & Neckers, L. M. The benzoquinone ansamycin 17-allylamino-17-demethoxygeldanamycin binds to HSP90 and shares important biologic activities with geldanamycin. Cancer Chemother. Pharmacol. 42, 273–279 (1998)

    Article  CAS  Google Scholar 

  2. Xu, W. et al. Sensitivity of mature Erbb2 to geldanamycin is conferred by its kinase domain and is mediated by the chaperone protein Hsp90. J. Biol. Chem. 276, 3702–3708 (2001)

    Article  CAS  Google Scholar 

  3. Sato, S., Fujita, N. & Tsuruo, T. Modulation of Akt kinase activity by binding to Hsp90. Proc. Natl Acad. Sci. USA 97, 10832–10837 (2000)

    Article  ADS  CAS  Google Scholar 

  4. Basso, A. D., Solit, D. B., Munster, P. N. & Rosen, N. Ansamycin antibiotics inhibit Akt activation and cyclin D expression in breast cancer cells that overexpress HER2. Oncogene 21, 1159–1166 (2002)

    Article  CAS  Google Scholar 

  5. Schulte, T. W., Blagosklonny, M. V., Ingui, C. & Neckers, L. Disruption of the Raf-1-Hsp90 molecular complex results in destabilization of Raf-1 and loss of Raf-1-Ras association. J. Biol. Chem. 270, 24585–24588 (1995)

    Article  CAS  Google Scholar 

  6. An, W. G., Schulte, T. W. & Neckers, L. M. The heat shock protein 90 antagonist geldanamycin alters chaperone association with p210bcr-abl and v-src proteins before their degradation by the proteasome. Cell Growth Differ. 11, 355–360 (2000)

    CAS  PubMed  Google Scholar 

  7. Blagosklonny, M. V., Toretsky, J. & Neckers, L. Geldanamycin selectively destabilizes and conformationally alters mutated p53. Oncogene 11, 933–939 (1995)

    CAS  PubMed  Google Scholar 

  8. Whitesell, L., Mimnaugh, E. G., De Costa, B., Myers, C. E. & Neckers, L. M. Inhibition of heat shock protein HSP90-pp60v-src heteroprotein complex formation by benzoquinone ansamycins: Essential role for stress proteins in oncogenic transformation. Proc. Natl Acad. Sci. USA 91, 8324–8328 (1994)

    Article  ADS  CAS  Google Scholar 

  9. Miller, P. et al. Depletion of the erbB-2 gene product p185 by benzoquinoid ansamycins. Cancer Res. 54, 2724–2730 (1994)

    CAS  PubMed  Google Scholar 

  10. Mimnaugh, E. G., Chavany, C. & Neckers, L. Polyubiquitination and proteasomal degradation of the p185c-erbB-2 receptor protein-tyrosine kinase induced by geldanamycin. J. Biol. Chem. 271, 22796–22801 (1996)

    Article  CAS  Google Scholar 

  11. Schulte, T. W., An, W. G. & Neckers, L. M. Geldanamycin-induced destabilization of Raf-1 involves the proteasome. Biochem. Biophys. Res. Commun. 239, 655–659 (1997)

    Article  CAS  Google Scholar 

  12. Workman, P. Auditing the pharmacological accounts for hsp90 molecular chaperone inhibitors: Unfolding the relationship between pharmacokinetics and pharmacodynamics. Mol. Cancer Ther. 2, 131–138 (2003)

    Article  CAS  Google Scholar 

  13. Isaacs, J. S., Xu, W. & Neckers, L. Heat shock protein 90 as a molecular target for cancer therapeutics. Cancer Cell 3, 213–217 (2003)

    Article  CAS  Google Scholar 

  14. Prodromou, C. et al. Identification and structural characterization of the ATP/ADP-binding site in the Hsp90 molecular chaperone. Cell 90, 65–75 (1997)

    Article  CAS  Google Scholar 

  15. Stebbins, C. E. et al. Crystal structure of an Hsp90-geldanamycin complex: Targeting of a protein chaperone by an antitumor agent. Cell 89, 239–250 (1997)

    Article  CAS  Google Scholar 

  16. Obermann, W. M., Sondermann, H., Russo, A. A., Pavletich, N. P. & Hartl, F. U. In vivo function of Hsp90 is dependent on ATP binding and ATP hydrolysis. J. Cell Biol. 143, 901–910 (1998)

    Article  CAS  Google Scholar 

  17. Panaretou, B. et al. ATP binding and hydrolysis are essential to the function of the Hsp90 molecular chaperone in vivo. EMBO J. 17, 4829–4836 (1998)

    Article  CAS  Google Scholar 

  18. Grenert, J. P., Johnson, B. D. & Toft, D. O. The importance of ATP binding and hydrolysis by hsp90 in formation and function of protein heterocomplexes. J. Biol. Chem. 274, 17525–17533 (1999)

    Article  CAS  Google Scholar 

  19. Solit, D. B. et al. 17-Allylamino-17-demethoxygeldanamycin induces the degradation of androgen receptor and HER-2/neu and inhibits the growth of prostate cancer xenografts. Clin. Cancer Res. 8, 986–993 (2002)

    ADS  CAS  PubMed  Google Scholar 

  20. Blagosklonny, M. V. Hsp-90-associated oncoproteins: Multiple targets of geldanamycin and its analogs. Leukemia 16, 455–462 (2002)

    Article  CAS  Google Scholar 

  21. Chiosis, G. et al. A small molecule designed to bind to the adenine nucleotide pocket of Hsp90 causes Her2 degradation and the growth arrest and differentiation of breast cancer cells. Chem. Biol. 8, 289–299 (2001)

    Article  CAS  Google Scholar 

  22. Egorin, M. J. et al. PC3 human prostate xenograft retention of, and oncoprotein modulation by 17-(allylamino)-17-demothoxygeldanamycin(17AAG) in vivo. Proc. Am. Assoc. Cancer Res. 40, 3409 (1999)

    Google Scholar 

  23. Chiosis, G. et al. 17AAG: Low target binding affinity and potent cell activity-finding an explanation. Mol. Cancer Ther. 2, 123–129 (2003)

    CAS  PubMed  Google Scholar 

  24. Richter, K. & Buchner, J. Hsp90: Chaperoning signal transduction. J. Cell. Physiol. 188, 281–290 (2001)

    Article  CAS  Google Scholar 

  25. Schuh, S. et al. A 90,000-dalton binding protein common to both steroid receptors and the Rous sarcoma virus transforming protein, pp60v-src. J. Biol. Chem. 260, 14292–14296 (1985)

    CAS  PubMed  Google Scholar 

  26. Kosano, H., Stensgard, B., Charlesworth, M. C., McMahon, N. & Toft, D. The assembly of progesterone receptor-hsp90 complexes using purified proteins. J. Biol. Chem. 273, 32973–32979 (1998)

    Article  CAS  Google Scholar 

  27. Gress, T. M. et al. Differential expression of heat shock proteins in pancreatic carcinoma. Cancer Res. 54, 547–551 (1994)

    CAS  PubMed  Google Scholar 

  28. Yano, M., Naito, Z., Tanaka, S. & Asano, G. Expression and roles of heat shock proteins in human breast cancer. Jpn J. Cancer Res. 87, 908–915 (1996)

    Article  CAS  Google Scholar 

  29. Rutherford, S. L. & Lindquist, S. Hsp90 as a capacitor for morphological evolution. Nature 396, 336–342 (1998)

    Article  ADS  CAS  Google Scholar 

  30. Queitsch, C., Sangster, T. A. & Lindquist, S. Hsp90 as a capacitor of phenotypic variation. Nature 417, 618–624 (2002)

    Article  ADS  CAS  Google Scholar 

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Acknowledgements

We thank D. Toft for critical reading of the manuscript. We also thank G. Timony for preparing the scatter plots, P. Karjian for quantitative western blotting of Hop and p23, and other team members at Conforma Therapeutics Corporation for discussions and suggestions.

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  1. Conforma Therapeutics Corporation, 9393 Towne Centre Drive, Suite 240, San Diego, California, 92121, USA

    Adeela Kamal, Lia Thao, John Sensintaffar, Lin Zhang, Marcus F. Boehm, Lawrence C. Fritz & Francis J. Burrows

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Correspondence to Francis J. Burrows.

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Kamal, A., Thao, L., Sensintaffar, J. et al. A high-affinity conformation of Hsp90 confers tumour selectivity on Hsp90 inhibitors. Nature 425, 407–410 (2003). https://doi.org/10.1038/nature01913

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