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Mechanism of folding and assembly of a small tetrameric protein domain from tumor suppressor p53

Nature Structural Biology volume 6, pages 191–198 (1999)Cite this article

Abstract

We have analyzed the folding pathway of the tetramerization domain of the tumor suppressor protein p53. Structures of transition states were determined from Φ–values for 25 mutations, including leucine to norvaline, and the analysis encompassed nearly every residue in the domain. Denatured monomers fold and dimerize, through a transition state with little native structure, to form a transient, highly structured dimeric intermediate. The intermediate dimerizes, through a native–like transition state with the primary dimers fully folded but with interdimer interactions only partially formed, to form the native tetramer as a 'dimer of dimers'.

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Figure 1: Ribbon model of the p53tet structure.
Figure 2: a, Kinetics of unfolding of p53tetS in 4 M GdmCl followed by CD.
Figure 3: a, Kinetics of refolding of GdmCl–denatured p53tetS followed by CD.
Figure 4: Variation in the rate of unfolding with GdmCl concentration for some representative p53tet tetrameric mutants.
Figure 5: Residues with fractional Φ–values in analysis of p53tet unfolding.
Figure 6: Proposed qualitative reaction coordinate diagram for the folding and unfolding of tetrameric wild type p53tet and most tetrameric mutants.

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Acknowledgements

We gratefully acknowledge M. Bycroft, C. Johnson and J.L. Neira for scientific discussions and expert advice. This work was supported by the CRC of the UK. M.G.M. was supported by a grant from the European Union.

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Author notes

  1. Mauricio G. Mateu and Alan R. Fersht: On leave from Universidad Autónoma de Madrid, Cantoblanco 28049 Madrid, Spain.

Authors and Affiliations

  1. Cambridge University Chemical Laboratory and Cambridge Centre for Protein Engineering, MRC Centre, Hills Road, Cambridge, CB2 2QH, UK

    Mauricio G. Mateu & Manuel M. Sánchez Del Pino

  2. Alan R. Fersht

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  1. Mauricio G. Mateu
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  2. Manuel M. Sánchez Del Pino
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  3. Alan R. Fersht
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Correspondence to Alan R. Fersht.

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Mateu, M., Sánchez Del Pino, M. & Fersht, A. Mechanism of folding and assembly of a small tetrameric protein domain from tumor suppressor p53. Nat Struct Mol Biol 6, 191–198 (1999). https://doi.org/10.1038/5880

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