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

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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

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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