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Crystal Structure of an Ancient Protein: Evolution by Conformational Epistasis

Science, 14 September 2007
Vol. 317, Issue 5844, p. 1544-1548
DOI: 10.1126/science.1142819

Crystal Structure of an Ancient Protein: Evolution by Conformational Epistasis

  1. Eric A. Ortlund1,*,
  2. Jamie T. Bridgham2,*,
  3. Matthew R. Redinbo1,
  4. Joseph W. Thornton2,
  1. 1 Department of Chemistry, University of North Carolina, Chapel Hill, NC 27599, USA.
  2. 2 Center for Ecology and Evolutionary Biology, University of Oregon, Eugene, OR 97403, USA.
  1. To whom correspondence should be addressed. E-mail: joet{at}
  • * These authors contributed equally to this work.


The structural mechanisms by which proteins have evolved new functions are known only indirectly. We report x-ray crystal structures of a resurrected ancestral protein—the ∼450 million-year-old precursor of vertebrate glucocorticoid (GR) and mineralocorticoid (MR) receptors. Using structural, phylogenetic, and functional analysis, we identify the specific set of historical mutations that recapitulate the evolution of GR's hormone specificity from an MR-like ancestor. These substitutions repositioned crucial residues to create new receptor-ligand and intraprotein contacts. Strong epistatic interactions occur because one substitution changes the conformational position of another site. “Permissive” mutations—substitutions of no immediate consequence, which stabilize specific elements of the protein and allow it to tolerate subsequent function-switching changes—played a major role in determining GR's evolutionary trajectory.

    • Received for publication 21 March 2007.
    • Accepted for publication 6 July 2007.


    E. A. Ortlund, J. T. Bridgham, M. R. Redinbo, and J. W. Thornton, Crystal Structure of an Ancient Protein: Evolution by Conformational Epistasis. Science 317, 1544-1548 (2007).

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