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Protein conformational stabilities can be determined from hydrogen exchange rates

Nature Structural Biology volume 6, pages 910–912 (1999)Cite this article

Abstract

Measuring protein conformational stability is one key to solving the protein folding problem. The conformational stability is the free energy change of the unfolding reaction, F ↔ U, under ambient conditions, ΔGU = GU - GF. Traditional methods of measuring ΔGU are solvent (urea or guanidinium chloride (GdmCl)) or thermal denaturation1. Solvent denaturation curves are generally analyzed using the linear extrapolation method (LEM): ΔG = ΔGU(H2O) - m[denaturant]  (1) where m is a measure of the dependence of ΔG on denaturant, and ΔGU(H2O) is an estimate of the conformational stability that assumes that the linear dependence of ΔG on denaturant observed in the transition region continues to 0 M denaturant. Thermal denaturation experiments yield the melting temperature, Tm, the enthalpy change at Tm, ΔHm, and the heat capacity change, ΔCp, which can then be used to calculate ΔGU at any temperature T, ΔGU(T), with the Gibbs–Helmholtz equation: ΔGU(T) = ΔHm(1 - T/Tm) + ΔCp[T - Tm - T ln (T/Tm)]  (2)

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Acknowledgements

The authors thank C. Woodward for a preprint of a recent review on hydrogen exchange, S. Marqusee and M. Llinas for sending unpublished data, and many colleagues for helpful discussions and comments. Support from National Institutes of Health and the Robert A. Welch Foundation is acknowledged. J.M.S. is an American Heart Association Established Investigator, and C.N.P. is supported by the Tom and Jean McMullin Professorship.

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Authors and Affiliations

  1. Departments of Medical Biochemistry and Genetics, and of Biochemistry and Biophysics, Center for Macromolecular Design, Texas A & M University, College Station, 77843, Texas, USA

    Beatrice M.P. Huyghues-Despointes, J. Martin Scholtz & C. Nick Pace

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  1. Beatrice M.P. Huyghues-Despointes
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  2. J. Martin Scholtz
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Correspondence to J. Martin Scholtz or C. Nick Pace.

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Huyghues-Despointes, B., Scholtz, J. & Pace, C. Protein conformational stabilities can be determined from hydrogen exchange rates. Nat Struct Mol Biol 6, 910–912 (1999). https://doi.org/10.1038/13273

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