Linking thermodynamics and measurements of protein stability

TL;DR

This paper reviews the thermodynamic principles and experimental methods for analyzing protein stability through equilibrium unfolding experiments, emphasizing the importance of modeling folding equilibria and baselines.

Contribution

It provides a comprehensive overview of the theoretical background, equations, and modeling approaches for interpreting protein unfolding data.

Findings

Highlights the significance of modeling folding equilibria.

Emphasizes the need for accurate baseline modeling.

Summarizes methods for extracting thermodynamic parameters.

Abstract

We review the background, theory and general equations for the analysis of equilibrium protein unfolding experiments, focusing on denaturant and heat-induced unfolding. The primary focus is on the thermodynamics of reversible folding/unfolding transitions and the experimental methods that are available for extracting thermodynamic parameters. We highlight the importance of modelling both how the folding equilibrium depends on a perturbing variable such as temperature or denaturant concentration, and the importance of modelling the baselines in the experimental observables.