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The energetics of T4 lysozyme reveal a hierarchy of conformations

Nature Structural Biology volume 6, pages 1072–1078 (1999)Cite this article

Abstract

We have used native state exchange to examine the energy landscape of the well-characterized protein T4 lysozyme. Although the protein exhibits two-state behavior by traditional probes, the energy landscape determined here is much more complex. The average stability of the C-terminal subdomain is significantly higher than that for the N-terminus suggesting at least two regions of unfolding. At a more detailed level, there appears to be a broad continuum of stabilities throughout each region. The overall subdomain hierarchy of energies does not mirror data on the folding pathway for this protein, challenging the relationship between energy landscapes and folding trajectories.

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Figure 1: Equilibrium unfolding of T4 lysozyme.
Figure 2: Distribution of unfolding free energies (ΔGunf) for T4 lysozyme.
Figure 3: The calculated unfolding free energies (ΔGunf) and corresponding m-values define a broad continuum of stabilities.
Figure 4: The measured free energy of exchange (ΔGunf) is correlated with the Cα–Cα distance from Ile 100 in the E-helix.

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Acknowledgements

We thank K. Fischer, I. Griswold, J. Hollien, G. Lazar, and M.J. Parker for discussion and critical reading of the manuscript. We also thank A.J. Wand and E. Fuentes for discussion about data analysis. This work was supported by grants from the National Institutes of Health (S.M. and F.W.D.) and the W. M. Keck Foundation. M.L. was supported by an NIH Molecular Biophysics Training Grant and B.G. was supported by a Department of Health and Human Services, National Research Service Award.

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Authors and Affiliations

  1. Department of Molecular and Cell Biology, 229 Stanley Hall, University of California, Berkeley, Berkeley, 94720, California, USA

    Manuel Llinás & Susan Marqusee

  2. Institute of Molecular Biology and Departments of Chemistry and Physics, University of Oregon, Eugene, 97403, Oregon, USA

    Blake Gillespie & Frederick W. Dahlquist

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  1. Manuel Llinás
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Correspondence to Susan Marqusee.

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Llinás, M., Gillespie, B., Dahlquist, F. et al. The energetics of T4 lysozyme reveal a hierarchy of conformations. Nat Struct Mol Biol 6, 1072–1078 (1999). https://doi.org/10.1038/14956

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