Protein stability modulated by a conformational effector: effects of trifluoroethanol on bovine serum albumin

Phys Chem Chem Phys. 2009 May 28;11(20):4007-18. doi: 10.1039/b818687a. Epub 2009 Mar 5.

Abstract

The link between the thermodynamic properties of a solution and the conformational space explored by a protein is of fundamental importance to understand and control solubility, misfolding and aggregation processes. Here, we study the thermodynamic and conformational stability of a model protein, bovine serum albumin (BSA), by addition of trifluoroethanol (TFE), which is known to affect both the solvent properties and the protein structure. The solvent-mediated pair-wise interactions are investigated by static and dynamic light scattering, and by small angle X-ray scattering. The protein conformational details are studied by far- and near-UV circular dichroism (CD), and steady state fluorescence from tryptophan and from 1-anilino-8-naphthalene sulfonate (ANS). At low TFE concentrations, our results show that protein-protein interaction is dominated by steric repulsion accompanied by a consistent protein solvation. Minor local conformational changes also occur, but they do not affect the stability of BSA. At TFE concentrations above the threshold of 16% v/v, attractive interactions become prevalent, along with conformational changes related to a loosening of BSA tertiary structure. The onset of thermodynamic instability is triggered by the enhancement of hydrophobic attraction over repulsion, due to minor local changes of protein conformation and hydration. In the present context, TFE acts as a conformational effector, since it affects the intermolecular interaction and the activity of the proteins in solution through a direct mechanism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anilino Naphthalenesulfonates / chemistry
  • Animals
  • Cattle
  • Circular Dichroism
  • Fluorescence
  • Light
  • Protein Conformation / drug effects
  • Protein Stability / drug effects
  • Scattering, Small Angle
  • Serum Albumin, Bovine / chemistry*
  • Thermodynamics
  • Trifluoroethanol / chemistry
  • Trifluoroethanol / pharmacology*
  • Ultraviolet Rays
  • X-Ray Diffraction

Substances

  • Anilino Naphthalenesulfonates
  • Serum Albumin, Bovine
  • 1-anilino-8-naphthalenesulfonate
  • Trifluoroethanol