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Substantial increase of protein stability by multiple disulphide bonds

Nature volume 342, pages 291–293 (1989)Cite this article

Abstract

DISULPHIDE bonds can significantly stabilize the native structures of proteins1–3. The effect is presumed to be due mainly to a decrease in the configurational chain entropy of the unfolded polypeptide4–7. In phage T4 lysozyme, a disulphide-free enzyme, engineered disulphide mutants that crosslink residues 3–97, 9–164 and 21–142 are significantly more stable than the wild-type protein8–11. To investigate the effect of multiple-disulphide bonds on protein stability, mutants were constructed in which two or three stabilizing disulphide bridges were combined in the same protein. Reversible thermal denaturation shows that the increase in melting temperature resulting from the individual disulphide bonds is approximately additive. The triple-disulphide variant unfolds at a temperature 23.4 °C higher than wild-type lysozyme. The results demonstrate that a combination of disulphide bonds, each of which contributes to stability, can achieve substantial overall improvement in the stability of a protein.

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Author notes

  1. Masazumi Matsumura

    Present address: Department of Immunology, Research Institute of Scripps Clinic, 10666 North Torrey Pines Road, La Jolla, California, 92037, USA

Authors and Affiliations

  1. Institute of Molecular Biology and Department of Physics, University of Oregon, Eugene, Oregon, 97403, USA

    Masazumi Matsumura, Giovanni Signor & Brian W. Matthews

Authors
  1. Masazumi Matsumura
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  2. Giovanni Signor
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  3. Brian W. Matthews
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Matsumura, M., Signor, G. & Matthews, B. Substantial increase of protein stability by multiple disulphide bonds. Nature 342, 291–293 (1989). https://doi.org/10.1038/342291a0

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