Abstract
Hsp90 has emerged as a key chemotherapeutic target for the development of drugs for the treatment of cancer and neurodegenerative diseases. The shortcomings of many of the Hsp90 inhibitors that have made it to clinical trials have bolstered the need to identify new lead compounds with superior properties. Here, we describe a high-throughput screen for the identification of Hsp90 inhibitors based on the refolding of thermally denatured firefly luciferase.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Whitesell L, Mimnaugh EG, De Costa B, Myers CE, Neckers LM (1994) 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 U S A 91:8324–8328
Hanahan D, Weinberg RA (2000) The hallmarks of cancer. Cell 100:57–70
Hanahan D, Weinberg RA (2011) Hallmarks of cancer: the next generation. Cell 144:646–674
Bishop SC, Burlison JA, Blagg BS (2007) Hsp90: a novel target for the disruption of multiple signaling cascades. Curr Cancer Drug Targets 7:369–388
Koga F, Kihara K, Neckers L (2009) Inhibition of cancer invasion and metastasis by targeting the molecular chaperone heat-shock protein 90. Anticancer Res 29:797–807
Grenert JP, Johnson BD, Toft DO (1999) The importance of ATP binding and hydrolysis by hsp90 in formation and function of protein heterocomplexes. J Biol Chem 274:17525–17533
Roe SM, Prodromou C, O’Brien R, Ladbury JE, Piper PW, Pearl LH (1999) Structural basis for inhibition of the Hsp90 molecular chaperone by the antitumor antibiotics radicicol and geldanamycin. J Med Chem 42:260–266
Neckers L, Workman P (2012) Hsp90 molecular chaperone inhibitors: are we there yet? Clin Cancer Res 18:64–76
Davenport J, Balch M, Galam L, Girgis A, Hall J, Blagg BS, Matts RL (2014) High-throughput screen of natural product libraries for hsp90 inhibitors. Biology (Basel) 3:101–138
Galam L, Hadden MK, Ma Z, Ye QZ, Yun BG, Blagg BS, Matts RL (2007) High-throughput assay for the identification of Hsp90 inhibitors based on Hsp90-dependent refolding of firefly luciferase. Bioorg Med Chem 15:1939–1946
Donnelly A, Blagg BS (2008) Novobiocin and additional inhibitors of the Hsp90 C-terminal nucleotide-binding pocket. Curr Med Chem 15:2702–2717
Thulasiraman V, Matts RL (1998) Luciferase renaturation assays of chaperones and chaperone antagonists. Methods Mol Biol 102:129–141
Thulasiraman V, Yun BG, Uma S, Gu Y, Scroggins BT, Matts RL (2002) Differential inhibition of Hsc70 activities by two Hsc70-binding peptides. Biochemistry 41:3742–3753
Uma S, Thulasiraman V, Matts RL (1999) Dual role for Hsc70 in the biogenesis and regulation of the heme-regulated kinase of the alpha subunit of eukaryotic translation initiation factor 2. Mol Cell Biol 19:5861–5871
Jackson RJ, Hunt T (1983) Preparation and use of nuclease-treated rabbit reticulocyte lysates for the translation of eukaryotic messenger RNA. Methods Enzymol 96:50–74
Matts RL, Hurst R (1992) The relationship between protein synthesis and heat shock proteins levels in rabbit reticulocyte lysates. J Biol Chem 267:18168–18174
Matts RL, Schatz JR, Hurst R, Kagen R (1991) Toxic heavy metal ions activate the heme-regulated eukaryotic initiation factor-2 alpha kinase by inhibiting the capacity of hemin-supplemented reticulocyte lysates to reduce disulfide bonds. J Biol Chem 266:12695–12702
Merrick WC (1983) Translation of exogenous mRNAs in reticulocyte lysates. Methods Enzymol 101:606–615
Schumacher RJ, Hurst R, Sullivan WP, McMahon NJ, Toft DO, Matts RL (1994) ATP-dependent chaperoning activity of reticulocyte lysate. J Biol Chem 269:9493–9499
Matts RL, Manjarrez JR (2009) Assays for identification of Hsp90 inhibitors and biochemical methods for discriminating their mechanism of action. Curr Top Med Chem 9:1462–1478
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Davenport, J., Galam, L., Matts, R.L. (2018). A High-Throughput Screen for Inhibitors of the Hsp90-Chaperone Machine. In: Calderwood, S., Prince, T. (eds) Chaperones. Methods in Molecular Biology, vol 1709. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7477-1_7
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7477-1_7
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7476-4
Online ISBN: 978-1-4939-7477-1
eBook Packages: Springer Protocols