Equilibrium Denaturation Experiment


Figure adopted from Grantcharova and Baker, Biochemistry, 1997, 36(50):15685

 
Equilibrium GdmCl denaturation followed by fluorescence (filled circle) and CD (filled triangle) at 295 K. Fluorescence was recorded at 225 nm and CD at 235 nm. Solid lines are the fits to the experimental data using the linear extrapolation model. Baselines were corrected for sloping before conversion to fraction folded.

signal = N0 + CntT + CngD

+[(D0+DntT+DngD)exp(-G/RT)]/[1+exp(-G/RT)] (1)

where

GU = Hg + mD - TSg Cp [Tg - T + T ln(T/Tg)] (2)

 

Figure adopted from Grantcharova and Baker, Biochemistry, 1997, 36(50):15685

Three-dimensional guanidine-temperature denaturation surface. Closed circles represent experimentally determined values from the individual temperature melts at different GdmCl concentrations. Continuous net is the best fit of eq 1 or 2 to the experimental data. The GU estimates from two models were similar (4.7 +/- 0.22 and 4.44 +/- 0.2 kcal/mol).

[Note] The 3D denaturation method reduces thhe uncertainties in estimations quanties in 0 M denaturant since the extroplations are based on more data compared to a GdmCl denaturation experiment at a single temperature.

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Danny S. Hsu 1999-2000. All rights reserved.