Single-shot NMR measurement of protein unfolding landscapes

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

    Research output: Contribution to journalArticle

    Abstract

    The transient unfolding events from the native state of a protein towards higher energy states can be closely investigated by studying the process of hydrogen exchange. Here, we present BLUU-Tramp (Biophysics Laboratory University of Udine - Temperature ramp), a new method to measure the rates for the exchange process and the underlying equilibrium thermodynamic parameters, using just a single sample preparation, in a single experiment that lasts some 20 to 60 h depending on the protein thermal stability, to record hundreds of points over a virtually continuous temperature window. The method is suitable also in presence of other proteins in the sample, if only the target protein is 15N-labelled. This allows the complete thermodynamic description of the unfolding landscape at an atomic level in the presence of small or macromolecular ligands or cosolutes, or in physiological environments. The method was successfully tested with human ubiquitin. Then the unfolding thermodynamic parameters were satisfactorily determined for the amyloidogenic protein β2-microglobulin, in aqueous buffer and in synovial liquid, that is the natural medium of amyloid deposition in joints.

    Original languageEnglish (US)
    Pages (from-to)842-849
    Number of pages8
    JournalBiochimica et Biophysica Acta - Proteins and Proteomics
    Volume1824
    Issue number6
    DOIs
    StatePublished - Jun 1 2012

    Fingerprint

    Protein Unfolding
    Thermodynamics
    Nuclear magnetic resonance
    Amyloidogenic Proteins
    Biophysics
    Architectural Accessibility
    Proteins
    Temperature
    Protein Stability
    Ubiquitin
    Amyloid
    Hydrogen
    Buffers
    Hot Temperature
    Joints
    Electron energy levels
    Ligands
    Thermodynamic stability
    Liquids
    Experiments

    Keywords

    • Energy landscape
    • H-D exchange
    • Protein NMR
    • Protein thermal denaturation
    • Protein unfolding
    • Unfolding thermodynamics

    ASJC Scopus subject areas

    • Biochemistry
    • Biophysics
    • Analytical Chemistry
    • Molecular Biology

    Cite this

    Rennella, E., Corazza, A., Codutti, L., Causero, A., Bellotti, V., Stoppini, M., ... Esposito, G. (2012). Single-shot NMR measurement of protein unfolding landscapes. Biochimica et Biophysica Acta - Proteins and Proteomics, 1824(6), 842-849. https://doi.org/10.1016/j.bbapap.2012.04.002

    Single-shot NMR measurement of protein unfolding landscapes. / Rennella, Enrico; Corazza, Alessandra; Codutti, Luca; Causero, Araldo; Bellotti, Vittorio; Stoppini, Monica; Viglino, Paolo; Fogolari, Federico; Esposito, Gennaro.

    In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1824, No. 6, 01.06.2012, p. 842-849.

    Research output: Contribution to journalArticle

    Rennella, E, Corazza, A, Codutti, L, Causero, A, Bellotti, V, Stoppini, M, Viglino, P, Fogolari, F & Esposito, G 2012, 'Single-shot NMR measurement of protein unfolding landscapes', Biochimica et Biophysica Acta - Proteins and Proteomics, vol. 1824, no. 6, pp. 842-849. https://doi.org/10.1016/j.bbapap.2012.04.002
    Rennella E, Corazza A, Codutti L, Causero A, Bellotti V, Stoppini M et al. Single-shot NMR measurement of protein unfolding landscapes. Biochimica et Biophysica Acta - Proteins and Proteomics. 2012 Jun 1;1824(6):842-849. https://doi.org/10.1016/j.bbapap.2012.04.002
    Rennella, Enrico ; Corazza, Alessandra ; Codutti, Luca ; Causero, Araldo ; Bellotti, Vittorio ; Stoppini, Monica ; Viglino, Paolo ; Fogolari, Federico ; Esposito, Gennaro. / Single-shot NMR measurement of protein unfolding landscapes. In: Biochimica et Biophysica Acta - Proteins and Proteomics. 2012 ; Vol. 1824, No. 6. pp. 842-849.
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