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Studying heat shock proteins through single-molecule mechanical manipulation

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Cell Stress and Chaperones Aims and scope

Abstract

Imbalances of cellular proteostasis are linked to ageing and human diseases, including neurodegenerative and neuromuscular diseases. Heat shock proteins (HSPs) and small heat shock proteins (sHSPs) together form a crucial core of the molecular chaperone family that plays a vital role in maintaining cellular proteostasis by shielding client proteins against aggregation and misfolding. sHSPs are thought to act as the first line of defence against protein unfolding/misfolding and have been suggested to act as “sponges” that rapidly sequester these aberrant species for further processing, refolding, or degradation, with the assistance of the HSP70 chaperone system. Understanding how these chaperones work at the molecular level will offer unprecedented insights for their manipulation as therapeutic avenues for the treatment of ageing and human disease. The evolution in single-molecule force spectroscopy techniques, such as optical tweezers (OT) and atomic force microscopy (AFM), over the last few decades have made it possible to explore at the single-molecule level the structural dynamics of HSPs and sHSPs and to examine the key molecular mechanisms underlying their chaperone activities. In this paper, we describe the working principles of OT and AFM and the experimental strategies used to employ these techniques to study molecular chaperones. We then describe the results of some of the most relevant single-molecule manipulation studies on HSPs and sHSPs and discuss how these findings suggest a more complex physiological role for these chaperones than previously assumed.

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Funding

S.C and C.C. acknowledge funding by Italian Ministry of University and Research (MIUR), Departments of excellence 2018–2022; E91I18001480001, PRIN—Progetti di Ricerca di Interesse Nazionale (2017 EX_ALS) and University of Modena and Reggio Emilia (FAR 2016).

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Authors and Affiliations

  1. Department of Physics, Informatics and Mathematics, University of Modena and Reggio Emilia, 41125, Modena, Italy

    Dhawal Choudhary & Ciro Cecconi

  2. Institute of Nanoscience S3, Consiglio Nazionale delle Ricerche, 41125, Modena, Italy

    Dhawal Choudhary & Ciro Cecconi

  3. Department of Biomedical, Metabolic and Neural Sciences, and Centre for Neuroscience and Neurotechnology, University of Modena and Reggio Emilia, via G. Campi 287, 41125, Modena, Italy

    Laura Mediani & Serena Carra

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  1. Dhawal Choudhary
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Correspondence to Serena Carra or Ciro Cecconi.

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Choudhary, D., Mediani, L., Carra, S. et al. Studying heat shock proteins through single-molecule mechanical manipulation. Cell Stress and Chaperones 25, 615–628 (2020). https://doi.org/10.1007/s12192-020-01096-y

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