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

ASJC Scopus subject areas

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

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    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, 134(10), 4457-4460. https://doi.org/10.1021/ja209004q