Professional Documents
Culture Documents
2011
Biomolecular Engineering, CP 165/64 Universit Libre de Bruxelles, 50 Av. F.D. Roosevelt, B1050 Bruxelles, Belgium
2Laboratoire
Belgium
Abstract: All biological phenomena depend on molecular recognition, which is either intermolecular like in
ligand binding to a macromolecule or intramolecular like in protein folding. As a result, understanding the
relationship between the structure of proteins and the energetics of their stability and binding with others
(bio)molecules is a very interesting point in biochemistry and biotechnology. It is essential to the engineering
of stable proteins and to the structure-based design of pharmaceutical ligands. The parameter generally used to
characterize the stability of a system (the folded and unfolded state of the protein for example) is the equilibrium
constant (K) or the free energy (G), which is the sum of enthalpic (H) and entropic (S) terms. These
parameters are temperature dependent through the heat capacity change (Cp). The thermodynamic parameters
H and Cp can be derived from spectroscopic experiments, using the vant Hoff method, or measured directly
using calorimetry. Along with isothermal titration calorimetry (ITC), differential scanning calorimetry (DSC) is
a powerful method, less described than ITC, for measuring directly the thermodynamic parameters which
charaterize biomolecules.
In this article, we summarize the principal thermodynamics parameters, describe the DSC approach and review
some systems to which it has been applied. DSC is much used for the study of the stability and the folding of
biomolecules, but it can also be applied in order to understand biomolecular interactions and can thus be an
interesting technique in the process of drug design.
- H(T)
R
1
T
S(T)
R
d ln K (T) - H(T)
=
d (1/T)
R
When a macromolecule changes its thermodynamic state
(e.g., unfolds), a heat capacity change (Cp) is observed.
This change is due to the fact that the heat required to raise
the temperature of a solution of unfolded protein is greater
than that required for a solution of folded protein. Heat
capacity changes are primarily due to restructuring of the
solvent molecules around the non polar sidechains exposed
to the solvent during the unfolding process [2]. Assuming a
constant temperature-independent Cp, G is described by:
2005 Bentham Science Publishers Ltd.
2012
Bruylants et al.
TR
Cp[(T - Tm) - T ln ( T )]
Tm
Cp ln ( T )
Tm
Fig. (1). DSC experiment for the two-state unfolding of a globular protein (Adapted from Fig. 1 of ref. [1]).
2013
2014
Bruylants et al.
H m ( 1
nR
Tm
the tryptophan synthase beta2 subunit (Pfbeta2) [54], betalactoglobulins A and B [55], HIV-1 gp41 and gp160 [56],
NAD(+) synthetase [57], DNA ligase [58], streptokinase
from Streptococcus equisimilis were also studied by DSC
[59,60].
Cp ( ln Tm
1
Tm0 ) nR
Tm0
[L]
Tm0 1)} 1
Tm
H (Tm) - [ H m +
2015
Cp(Tm - Tm0)]
HB - T SB
2016
Bruylants et al.
Fig. (5). Increased thermal stability of ZAG upon ligand binding. Normalized DSC data, corrected for buffer base line showing
cooperative endothermic unfolding of wild-type (A) and recombinant ZAG (B) in the presence and absence of 0.2 mM DAUDA.
(Adapted from Fig. 4 of ref. [72]).
a)
2017
b)
Fig. (6). a) Schematic arrangement of subunits of the pyruvate dehydrogenase multienzyme complex: E1=pyruvate decarboxylase,
E2=dihydrolipoyl acetyltransferase, E3=dihydrolipoyl dehydrogenase, PSBD=peripheral subunit-binding domain, LD=lipoyl
domain, DD= recombinant di-domain representing the LD and PSBD of E2; b) DSC trace of the thermal denaturation of E3 and E1 in
the absence (dotted line) and presence (solid line) of PSBD (Adapted from Fig. 5 of ref. [77]).
2018
Bruylants et al.
1 = R ln[(1 + KB[L])
Tm
H
1
n
2019
5. CONCLUSION
[22]
[23]
[24]
[25]
2020
[26]
[27]
[28]
[29]
[30]
[31]
[32]
[33]
[34]
[35]
[36]
[37]
[38]
[39]
[40]
[41]
[42]
[43]
[44]
[45]
[46]
[47]
[48]
[49]
[50]
[51]
[52]
[53]
[54]
[55]
[56]
[57]
[58]
[59]
Bruylants et al.
[60]
[61]
[62]
[63]
[64]
[65]
[66]
[67]
[68]
[69]
[70]
[71]
[72]
[73]
[74]
[75]
[76]
[77]
[78]
[79]
[80]
[81]
[82]
[83]
[84]
[85]
[86]
[87]
[88]
[89]
[90]
[91]
[92]
[93]
[94]
[95]