Abstract
Stress-inducible heat-shock proteins (HSPs, like HSP70 and HSP27) are molecular chaperones that protect cells from stress damage by keeping cellular proteins in a folding competent state and preventing them from irreversible aggregation. HSP27 and HSP70 chaperone activities are useful indicators to test chemical products and physical stress impact on protein denaturation, to select HSP inhibitors, or to determine the implication of the chaperone function in other HSP activities, such as apoptosis. We have developed two simple and fast chaperone activity tests for HSP27 and HSP70 that we initially set up to test the effect of potential HSP inhibitors obtained after screening of chemical and small molecule libraries. These chaperone quantification tests are based on the capacity of HSP to counteract chemical or thermal protein aggregation.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Laskey, R. A., Honda, B. M., Mills, A. D., Morris, N. R., Wyllie, A. H., Mertz, J. E., De Roberts, E. M. and Gurdon, J. B. (1978). Chromatin assembly and transcription in eggs and oocytes of Xenopus laevis. Cold Spring Harb Symp Quant Biol. 42, 171–178.
Beckmann, R.P., Mizzen, L.E., Welch, W. J. (1990) Interaction of Hsp 70 with newly synthesized proteins: implications for protein folding and assembly. Science. 248, 850–854.
De Los Rios, P., Ben-Zvi, A., Slutsky, O., Azem, A., Goloubinoff, P. (2006) Hsp70 chaperones accelerate protein translocation and the unfolding of stable protein aggregates by entropic pulling. Proc Natl Acad Sci. USA. 103, 6166–6171.
Shi, Y., Thomas, J. O. (1992) The transport of proteins into the nucleus requires the 70-kilodalton heat shock protein or its cytosolic cognate. Mol Cell Biol. 12, 2186–2192.
Nollen, E. A., Brunsting, J. F., Roelofsen, H., Weber, L. A., Kampinga, H. H. (1999) In vivo chaperone activity of heat shock protein 70 and thermotolerance. Mol Cell Biol. 19, 2069–2079.
Ehrnsperger, M., Gräber, S., Gaestel, M., Buchner, J. (1997) Binding of non-native protein to Hsp25 during heat shock creates a reservoir of folding intermediates for reactivation. EMBO J. 16, 221–229.
Shashidharamurthy, R., Koteiche, H. A., Dong, J., McHaourab, H. S. (2005) Mechanism of chaperone function in small heat shock proteins: dissociation of the HSP27 oligomer is required for recognition and binding of destabilized T4 lysozyme. J Biol Chem. 280, 5281–5289.
Bruey, J. M., Paul, C., Fromentin, A., Hilpert, S., Arrigo, A. P., Solary, E., Garrido, C. (2000) Differential regulation of HSP27 oligomerization in tumor cells grown in vitro and in vivo. Oncogene. 19, 4855–4863.
Lelj-Garolla, B., Mauk, A. G. (2006) Self-association and chaperone activity of Hsp27 are thermally activated. J Biol Chem. 281, 8169–8174.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Gobbo, J., Gaucher-Di-Stasio, C., Weidmann, S., Guzzo, J., Garrido, C. (2011). Quantification of HSP27 and HSP70 Molecular Chaperone Activities. 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_11
Download citation
DOI: https://doi.org/10.1007/978-1-61779-295-3_11
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-294-6
Online ISBN: 978-1-61779-295-3
eBook Packages: Springer Protocols