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Defining the Role of Salt Bridges in Protein Stability

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Protein Structure, Stability, and Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 490))

Abstract

Although the energetic balance of forces stabilizing proteins has been established qualitatively over the last decades, quantification of the energetic contribution of particular interactions still poses serious problems. The reasons are the strong cooperativity and the interdependence of noncovalent interactions. Salt bridges are a typical example. One expects that ionizable side chains frequently form ion pairs in innumerable crystal structures. Since electrostatic attraction between opposite charges is strong per se, salt bridges can intuitively be regarded as an important factor stabilizing the native structure. Is that really so? In this chapter we critically reassess the available methods to delineate the role of electrostatic interactions and salt bridges to protein stability, and discuss the progress and the obstacles in this endeavor. The basic problem is that formation of salt bridges depends on the ionization properties of the participating groups, which is significantly influenced by the protein environment. Furthermore, salt bridges experience thermal fluctuations, continuously break and re-form, and their lifespan in solution is governed by the flexibility of the protein. Finally, electrostatic interactions are long-range and might be significant in the unfolded state, thus seriously influencing the energetic profile. Elimination of salt bridges by protonation/deprotonation at extreme pH or by mutation provides only rough energetic estimates, since there is no way to account for the nonadditive response of the protein moiety. From what we know so far, the strength of electrostatic interactions is strongly context-dependent, yet it is unlikely that salt bridges are dominant factors governing protein stability. Nevertheless, proteins from thermophiles and hyperthermophiles exhibit more, and frequently networked, salt bridges than proteins from the mesophilic counterparts. Increasing the thermal (not the thermodynamic) stability of proteins by optimization of charge–charge interactions is a good example for an evolutionary solution utilizing physical factors.

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Acknowledgments

Work in author’s own laboratories has been supported in part by the Swiss National Science Foundation (IJ) and by grant BIO4CT970129 of IV Biotechnology Program of the European Community (AK).

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  1. Biochemisches Institut der Universität Zürich, Zürich, Switzerland

    Ilian Jelesarov

  2. Department of Biosciences and Nutrition, Karolinska Institute, Stockholm, Sweden

    Andrey Karshikoff

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    Jelesarov, I., Karshikoff, A. (2009). Defining the Role of Salt Bridges in Protein Stability. In: Shriver, J. (eds) Protein Structure, Stability, and Interactions. Methods in Molecular Biology, vol 490. Humana Press. https://doi.org/10.1007/978-1-59745-367-7_10

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    • DOI: https://doi.org/10.1007/978-1-59745-367-7_10

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    • Publisher Name: Humana Press

    • Print ISBN: 978-1-58829-954-3

    • Online ISBN: 978-1-59745-367-7

    • eBook Packages: Springer Protocols

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