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Irreversible gelation of thermally unfolded proteins: structural and mechanical properties of lysozyme aggregates

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Abstract

The formation of protein aggregates is important in many fields of life science and technology. The morphological and mechanical properties of protein solutions depend upon the molecular conformation and thermodynamic and environmental conditions. Non-native or unfolded proteins may be kinetically trapped into irreversible aggregates and undergo precipitation or gelation. Here, we study the thermal aggregation of lysozyme in neutral solutions. We characterise the irreversible unfolding of lysozyme by differential scanning calorimetry. The structural properties of aggregates and their mechanisms of formation with the eventual gelation are studied at high temperature by spectroscopic, rheological and scattering techniques. The experiments show that irreversible micron-sized aggregates are organised into larger clusters according to a classical mechanism of diffusion and coagulation, which leads to a percolative transition at high concentrations. At a smaller length scale, optical and atomic force microscopy images reveal the existence of compact aggregates, which are the origin of the aggregation irreversibility.

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Abbreviations

SALS:

Small-angle light scattering

DSC:

Differential scanning calorimetry

AFM:

Atomic force microscopy

OD:

Optical density

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Acknowledgments

We thank R. Carrotta, G. Bellavia, A. Cupane, A. Emanuele, M. Leone, R. Noto and V. Vetri for relevant discussions and collaborations. We thank V. Foderà for the help in the use of the AFM instruments. This work was partially supported by the Italian National Research Council through the project “Intermolecular interaction in protein metastable solution”.

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Authors and Affiliations

  1. Institute of Biophysics at Palermo, Italian National Research Council, via U. La Malfa 153, 90146, Palermo, Italy

    Samuele Raccosta, Mauro Manno, Donatella Bulone, Daniela Giacomazza, Valeria Militello, Vincenzo Martorana & Pier Luigi San Biagio

  2. Department of Physical and Astronomical Sciences, University of Palermo, via Archirafi 36, 90133, Palermo, Italy

    Samuele Raccosta & Valeria Militello

Authors
  1. Samuele Raccosta
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  2. Mauro Manno
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  3. Donatella Bulone
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  4. Daniela Giacomazza
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  5. Valeria Militello
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  6. Vincenzo Martorana
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  7. Pier Luigi San Biagio
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Correspondence to Mauro Manno.

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Proceedings of the XIX Congress of the Italian Society of Pure and Applied Biophysics (SIBPA), Rome, September 2008.

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Raccosta, S., Manno, M., Bulone, D. et al. Irreversible gelation of thermally unfolded proteins: structural and mechanical properties of lysozyme aggregates. Eur Biophys J 39, 1007–1017 (2010). https://doi.org/10.1007/s00249-009-0503-4

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  • DOI: https://doi.org/10.1007/s00249-009-0503-4

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