Abstract
A 108 amino acid protein was designed and constructed from a reduced alphabet of seven amino acids. The 2.9 Å resolution X-ray crystal structure confirms that the protein is a four helix bundle, as it was designed to be. Hydrogen/deuterium exchange experiments reveal buried amide protons with protection factors in excess of 1 × 106 in the range characteristic of well protected protons in functional folded proteins (103–108) rather than protons in rapid exchange (0–102). The protein is monomeric at 1 mM, the concentration at which the exchange experiments were undertaken, indicating that the exchange factors are due to a unique stable tertiary structure fold, and not due to any higher order quaternary structure. Thermodynamic analysis provides an estimate of the free energy of folding of −9.3 kcal mole−1 at 25 °C, consistent with the free energy of folding derived from the protection factors of the most protected protons, indicating that global unfolding is required for exchange of the most protected protons.
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Schafmeister, C., LaPorte, S., Miercke, L. et al. A designed four helix bundle protein with native-like structure. Nat Struct Mol Biol 4, 1039–1046 (1997). https://doi.org/10.1038/nsb1297-1039
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DOI: https://doi.org/10.1038/nsb1297-1039
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