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The secondary structure and the thermal unfolding parameters of the S-layer protein from Lactobacillus salivarius

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Abstract

Surface layer (S-layer) proteins have been identified in the cell envelope of many organisms, such as bacteria and archaea. They self-assemble, forming monomolecular crystalline arrays. Isolated S-layer proteins are able to recrystallize into regular lattices, which proved useful in biotechnology. Here we investigate the structure and thermal unfolding of the S-layer protein isolated from Lactobacillus salivarius 16 strain of human origin. Using circular dichroism (CD) spectroscopy, and the software CDSSTR from DICHROWEB, CONTINLL from CDPro, as well as CDNN, we assess the fractions of the protein’s secondary structural elements at temperatures ranging between 10 and 90 °C, and predict the tertiary class of the protein. To study the thermal unfolding of the protein, we analyze the temperature dependence of the CD signal in the far- and near-UV domains. Fitting the experimental data by two- and three-state models of thermal unfolding, we infer the midpoint temperatures, the temperature dependence of the changes in Gibbs free energy, enthalpy, and entropy of the unfolding transitions in standard conditions, and the temperature dependence of the equilibrium constant. We also estimate the changes in heat capacity at constant pressure in standard conditions. The results indicate that the thermal unfolding of the S-layer protein from L. salivarius is highly cooperative, since changes in the secondary and tertiary structures occur simultaneously. The thermodynamic analysis predicts a “cold” transition, at about −3 °C, of both the secondary and tertiary structures. Our findings may be important for the use of S-layer proteins in biotechnology and in biomedical applications.

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Acknowledgments

This work was partially supported by a fellowship awarded to L.L. by the Federation of European Biochemical Societies (FEBS). We are grateful to the late Dr. Constantin Crăescu for guidance and hospitality at the Curie Institute INSERM U759, Paris, France (L.L).

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Authors and Affiliations

  1. Faculty of Physics, West University of Timisoara, Timisoara, Romania

    Liliana Lighezan

  2. The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, Sofia, Bulgaria

    Ralitsa Georgieva

  3. Center for Modeling Biological Systems and Data Analysis, Department of Functional Sciences, Victor Babes University of Medicine and Pharmacy Timisoara, Piata E. Murgu No. 2-4, 300041, Timisoara, Romania

    Adrian Neagu

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  1. Liliana Lighezan
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  2. Ralitsa Georgieva
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Correspondence to Adrian Neagu.

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Lighezan, L., Georgieva, R. & Neagu, A. The secondary structure and the thermal unfolding parameters of the S-layer protein from Lactobacillus salivarius . Eur Biophys J 45, 491–509 (2016). https://doi.org/10.1007/s00249-016-1117-2

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  • DOI: https://doi.org/10.1007/s00249-016-1117-2

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