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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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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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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DOI: https://doi.org/10.1038/75151
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