Uncoupler

Uncoupler analyzes coupled folding and binding equilibria in protein domains by performing global analysis of temperature-dependent isothermal titration calorimetry (ITC) and circular dichroism (CD) data to dissect thermodynamic contributions.

Key Features:

• Global analysis: Simultaneously examines folding and binding events across datasets to resolve coupled equilibria.
• Temperature dependence integration: Incorporates temperature-dependent ITC and CD measurements to characterize how folding and binding vary with temperature.
• Nested Gibbs–Helmholtz model: Applies a nested Gibbs-Helmholtz model to deconvolute intrinsic folding and binding contributions and determine enthalpic and entropic components.
• Non-linear enthalpy analysis: Handles non-linear relationships between binding enthalpy and temperature, exemplified in analyses of TPR domain interactions with Hsp90-derived peptides.

Scientific Applications:

• TPR domain thermodynamics: Dissects folding–binding coupling in tetratricopeptide repeat (TPR) domains and their interactions with Hsp90-derived peptides.
• Coupled folding–binding thermodynamics: Quantifies how coupled folding and binding affect observed affinities and the balance of enthalpic and entropic contributions in protein–protein recognition modules and signaling-related interactions.

Methodology:

Global analysis of temperature-dependent ITC and CD data; application of a nested Gibbs-Helmholtz model to separate and quantify folding and binding thermodynamic contributions; handling of non-linear enthalpy versus temperature relationships and evaluation of temperature-dependent effects on observed affinities.