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Two exposed amino acid residues confer thermostability on a cold shock protein

Nature Structural Biology volume 7, pages 380–383 (2000)Cite this article

Abstract

Thermophilic organisms produce proteins of exceptional stability. To understand protein thermostability at the molecular level we studied a pair of cold shock proteins, one of mesophilic and one of thermophilic origin, by systematic mutagenesis. Although the two proteins differ in sequence at 12 positions, two surface-exposed residues are responsible for the increase in stability of the thermophilic protein (by 15.8 kJ mol−1 at 70 °C). 11.5 kJ mol−1 originate from a predominantly electrostatic contribution of Arg 3 and 5.2 kJ mol−1 from hydrophobic interactions of Leu 66 at the carboxy terminus. The mesophilic protein could be converted to a highly thermostable form by changing the Glu residues at positions 3 and 66 to Arg and Leu, respectively. The variation of surface residues may thus provide a simple and powerful approach for increasing the thermostability of a protein.

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Figure 1: The mesophilic cold shock protein Bs-CspB differs from its thermophilic homolog, Bc-Csp, at 12 sequence positions.
Figure 2: Thermal stability curves.
Figure 3: Effect of all individual sequence differences on the stability of Bc-Csp.
Figure 4: The residues that contribute most to the difference in thermal stability between Bs-CspB and Bc-Csp are located near each other at the protein surface.

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Acknowledgements

We thank the members of our laboratories and the Marahiel laboratory for help and discussions as well as C. Brooks III (Scripps Research Institute) and C. Nick Pace (Texas A&M University) for a fruitful exchange of ideas about the electrostatic stabilization of proteins. This work was supported by grants from the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie.

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Authors and Affiliations

  1. Biochemisches Laboratorium, Universität Bayreuth , Bayreuth, D-95440, Germany

    Dieter Perl & Franz X. Schmid

  2. Forschungsgruppe Kristallographie, Max-Delbrück-Centrum für Molekulare Medizin, Robert-Roessle-Str. 10, Berlin, D-13125, Germany

    Uwe Mueller & Udo Heinemann

  3. Institut für Chemie – Kristallographie, Freie Universität Berlin, Takustr. 6, Berlin, D-14195 , Germany

    Uwe Mueller & Udo Heinemann

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  1. Dieter Perl
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Correspondence to Franz X. Schmid.

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Perl, D., Mueller, U., Heinemann, U. et al. Two exposed amino acid residues confer thermostability on a cold shock protein. Nat Struct Mol Biol 7, 380–383 (2000). https://doi.org/10.1038/75151

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