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Designing a 20-residue protein

Nature Structural Biology volume 9, pages 425–430 (2002)Cite this article

Abstract

Truncation and mutation of a poorly folded 39-residue peptide has produced 20-residue constructs that are >95% folded in water at physiological pH. These constructs optimize a novel fold, designated as the 'Trp-cage' motif, and are significantly more stable than any other miniprotein reported to date. Folding is cooperative and hydrophobically driven by the encapsulation of a Trp side chain in a sheath of Pro rings. As the smallest protein-like construct, Trp-cage miniproteins should provide a testing ground for both experimental studies and computational simulations of protein folding and unfolding pathways. Pro–Trp interactions may be a particularly effective strategy for the a priori design of self-folding peptides.

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Figure 1: Truncation and optimization of the C-terminal fold of EX4.
Figure 2: Folding measures for Trp-cage construct 5b.
Figure 3: NMR spectra and the structure derived for TC5b.

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Acknowledgements

Initial support came from a feasibility grant from the University of Washington Royalty Research Fund with continuing support from an NIH grant. We thank L. Serrano (EMBL-Heidelberg) for reminding us of the pH dependence of the helix-favoring QXXXD interaction.

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  1. Department of Chemistry, University of Washington, Seattle, 98195, Washington, USA

    Jonathan W. Neidigh, R. Matthew Fesinmeyer & Niels H. Andersen

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  1. Jonathan W. Neidigh
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  2. R. Matthew Fesinmeyer
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  3. Niels H. Andersen
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Correspondence to Niels H. Andersen.

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Neidigh, J., Fesinmeyer, R. & Andersen, N. Designing a 20-residue protein. Nat Struct Mol Biol 9, 425–430 (2002). https://doi.org/10.1038/nsb798

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