The Existence of Metastable Intermediate Lysozyme Conformation Highlights the Role of Alcohols in Altering Protein Stability

Marco Cannas, Michele D'Amico, Mauro Manno, Samuele Raccosta, Michele D'Amico, Vincenzo Martorana, Samuele Raccosta

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13 Citations (Scopus)

Abstract

Alcohols have a manifold effect on the conformational and thermodynamic stability of native proteins. Here, we study the effect of moderate concentrations of trifluoroethanol (TFE) on the thermal stability of hen egg-white lysozyme (HEWL), by far-UV circular dichroism and by steady-state and time-resolved photoluminescence of intrinsic tryptophans. Our results highlight that TFE affects lysozyme stability by preferential solvation of the protein molecule. Furthermore, we discovered the existence at 20% TFE of an equilibrium partially folded state of lysozyme, intermediate between the native and the unfolded state. A three-state model is therefore used to interpolate the thermal denaturation data. Our analysis explains how the stabilization of the intermediate conformation enhances the entropic contribution to unfolding, and thus decreases the unfolding temperature, while, at the same time, TFE enhances the conformational stability of the native fold at room temperature. Eventually, we challenged the ability of these intermediate structures to form supramolecular aggregates by heating experiments at different TFE concentrations.
Original languageEnglish
Pages (from-to)4078-4087
Number of pages10
JournalJOURNAL OF PHYSICAL CHEMISTRY. B, CONDENSED MATTER, MATERIALS, SURFACES, INTERFACES & BIOPHYSICAL
Volume115
Publication statusPublished - 2011

All Science Journal Classification (ASJC) codes

  • Physical and Theoretical Chemistry
  • Surfaces, Coatings and Films
  • Materials Chemistry

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