The temperature dependence of H/D exchange rates of proteins conveys information on the folding/unfolding thermodynamics and kinetics. Rather than running exchange only as a function of time at a single temperature, we run a two-variable experiment in which the time elapsing is coupled to a linear temperature ramp. This experiment is named BLUU-Tramp (Biophysics Laboratory Udine University Temperature ramp) From the unfolding ΔG values of each single backbone amide of β2-m (panel a), the structural determinants can be identified (panel b) .

A residue-by-residue complete thermodynamic analysis can be performed as shown below

Correlation among the thermodynamic parameters extracted from BLUU-Tramp measurements on human lysozyme