Determining the energy landscape of proteins by a fast isotope exchange NMR approach

Enrico Rennella, Alessandra Corazza, Luca Codutti, Vittorio Bellotti, Monica Stoppini, Paolo Viglino, Federico Fogolari, Gennaro Esposito

    Research output: Contribution to journalArticle

    Abstract

    We present a new and efficient NMR method, BLUU-Tramp (Biophysics Laboratory University of Udine temperature ramp), for the collection of hydrogen-deuterium exchange experiments as a function of time and temperature for small and medium-sized proteins. Exchange rates can be determined to extract the underlying thermodynamic equilibrium or kinetic parameters by sampling hundreds of points over a virtually continuous temperature ramp. Data are acquired in a single experimental session that lasts some 20-60 h, depending on the thermal stability of the protein. Subsequent analysis provides a complete thermodynamic description of the protein energy landscape. The global thermal unfolding process and the partial or local structure opening events can be fully determined at the single-residue resolution level. The proposed approach is shown to work successfully with the amyloidogenic protein β 2- microglobulin. With 15N-labeling, the unfolding landscape of a protein can also be studied in the presence of other unlabeled proteins and, in general, with ligands or cosolutes or in physiological environments.

    Original languageEnglish (US)
    Pages (from-to)4457-4460
    Number of pages4
    JournalJournal of the American Chemical Society
    Volume134
    Issue number10
    DOIs
    StatePublished - Mar 14 2012

    Fingerprint

    Isotopes
    Nuclear magnetic resonance
    Proteins
    Architectural Accessibility
    Thermodynamics
    Temperature
    Hot Temperature
    Amyloidogenic Proteins
    Biophysics
    Protein Stability
    Deuterium
    Hydrogen
    Kinetic parameters
    Labeling
    Ligands
    Thermodynamic stability
    Sampling
    Experiments

    ASJC Scopus subject areas

    • Chemistry(all)
    • Catalysis
    • Biochemistry
    • Colloid and Surface Chemistry

    Cite this

    Rennella, E., Corazza, A., Codutti, L., Bellotti, V., Stoppini, M., Viglino, P., ... Esposito, G. (2012). Determining the energy landscape of proteins by a fast isotope exchange NMR approach. Journal of the American Chemical Society, 134(10), 4457-4460. https://doi.org/10.1021/ja209004q

    Determining the energy landscape of proteins by a fast isotope exchange NMR approach. / Rennella, Enrico; Corazza, Alessandra; Codutti, Luca; Bellotti, Vittorio; Stoppini, Monica; Viglino, Paolo; Fogolari, Federico; Esposito, Gennaro.

    In: Journal of the American Chemical Society, Vol. 134, No. 10, 14.03.2012, p. 4457-4460.

    Research output: Contribution to journalArticle

    Rennella, E, Corazza, A, Codutti, L, Bellotti, V, Stoppini, M, Viglino, P, Fogolari, F & Esposito, G 2012, 'Determining the energy landscape of proteins by a fast isotope exchange NMR approach', Journal of the American Chemical Society, vol. 134, no. 10, pp. 4457-4460. https://doi.org/10.1021/ja209004q
    Rennella E, Corazza A, Codutti L, Bellotti V, Stoppini M, Viglino P et al. Determining the energy landscape of proteins by a fast isotope exchange NMR approach. Journal of the American Chemical Society. 2012 Mar 14;134(10):4457-4460. https://doi.org/10.1021/ja209004q
    Rennella, Enrico ; Corazza, Alessandra ; Codutti, Luca ; Bellotti, Vittorio ; Stoppini, Monica ; Viglino, Paolo ; Fogolari, Federico ; Esposito, Gennaro. / Determining the energy landscape of proteins by a fast isotope exchange NMR approach. In: Journal of the American Chemical Society. 2012 ; Vol. 134, No. 10. pp. 4457-4460.
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