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 journalArticlepeer-review


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
Issue number10
StatePublished - Mar 14 2012

ASJC Scopus subject areas

  • Catalysis
  • General Chemistry
  • Biochemistry
  • Colloid and Surface Chemistry


Dive into the research topics of 'Determining the energy landscape of proteins by a fast isotope exchange NMR approach'. Together they form a unique fingerprint.

Cite this