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The folding transition state between SH3 domains is conformationally restricted and evolutionarily conserved

Nature Structural Biology volume 6, pages 1010–1016 (1999)Cite this article

Abstract

The protein engineering analysis of the α-spectrin SH3 domain at three different stability conditions (pH 7.0, 3.5 and 2.5) reveals a folding transition state structured around the distal loop β-hairpin and the 310-helix. This region is impervious to overall changes in protein stability, suggesting a transition state ensemble with little conformational variability. Comparison with the Src SH3 domain (36% sequence homology) indicates that the transition state in this protein family may be conserved. Discrepancies at some positions can be rationalized in terms of the different interactions made by the different side chains in both domains. Brønsted plot analysis confirms the φ‡-U results and shows two folding subdomains for this small protein. These results, together with previous data on circular permutants of the α-spectrin SH3 domain, indicate that polypeptide topology and chain connectivity play a major role in the folding reaction of this protein family.

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Figure 1: Sequence and structure comparison of the Src and α-spectrin SH3 domains showing the residues mutated in both proteins.
Figure 2: Analysis of the α-spectrin SH3 domain transition state.
Figure 3: Comparison between the φ‡-U values for equivalent mutations in the WT α-spectrin at pH 3.

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Acknowledgements

J.C.M. acknowledges European Union for financial support by a postdoctoral TMR fellowship. This project has been partly financed by an EU network grant.

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  1. EMBL, Meyerhofstrasse 1, Heidelberg, 69117, Germany

    Jose C. Martínez & Luis Serrano

  2. Departamento de Química Física, Universidad de Granada, Granada, 18071-, Spain

    Jose C. Martínez

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  1. Jose C. Martínez
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  2. Luis Serrano
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Correspondence to Luis Serrano.

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Martínez, J., Serrano, L. The folding transition state between SH3 domains is conformationally restricted and evolutionarily conserved. Nat Struct Mol Biol 6, 1010–1016 (1999). https://doi.org/10.1038/14896

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