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Protein Heat Capacity: An Anomaly that Maybe Never Was

Alan Cooper*
School of Chemistry, College of Science and Engineering, Joseph Black Building, University of Glasgow, Glasgow G12 8QQ, U.K.

ABSTRACT Protein unfolding in aqueous solution is usually accompanied by an

increase in heat capacity (ΔCp), and this has long been regarded as somewhat
anomalous. However, neither the absolute heat capacities (Cp) of folded globular
proteins nor the heat capacity increments upon unfolding (ΔCp) are unusual in
comparison to values observed for order-disorder (melting) transitions in other
organic substances. The consequences for protein stability, including cold dena-
turation, enthalpy-entropy compensation, and the temperature of maximum
stability, may seem counterintuitive but should not be unexpected. Nevertheless,
while perhaps not so anomalous as once thought, quantitative interpretation of
protein heat capacity and related effects remains a theoretical challenge.

A
round 1757, Joseph Black, a colleague from an earlier
era in Glasgow, was puzzled by his observations that
certain materials took longer to warm up than others.
Ice, water, and various “spiritous liquors” were particularly
intriguing, and it is said that “He waited with impatience for
the winter” so that he could continue his investigations on
heating and cooling.1 (There was no ice house in Glasgow, and
refrigeration had not yet been invented, though his professor
and mentor, William Cullen, was working on it.) This was no
idle pursuit; James Watt, Black's friend and “mathematical
instrument maker to the University”, at that time was con-
cerned with improving the efficiency of steam engines. What
Black showed was that substances undergoing heat-induced
phase transitions ; melting, boiling, and evaporation ; did
so without an increase in temperature. This led to the concept
of “latent” (i.e., hidden) heat, what we now call heat capacity,
in which heat energy is somehow used to do something other
than simply raise temperature.
Heat Capacity ; The Basics. As the name implies, heat
capacity is a measure of the capacity of any object to take up
heat energy. At the molecular level, this heat energy will be
distributed among the available degrees of freedom and
partitioned into kinetic energies (related to vibrational, rota-
tional, or translational motions) or potential energies (related
to changes in interatomic potential energies, bond stretching,
bending, breaking, and so forth). It follows that the more ways
there are of distributing heat energy in a substance, the higher
will be its heat capacity. This roughly explains why liquid water
has such a relatively high heat capacity; heat energy is diverted
into breaking residual intermolecular hydrogen bonds rather
than raising the temperature.
Heat capacity is central to fundamental thermodynamics
because both the absolute enthalpy (H) and the entropy (S) of
any object are integral functions of its heat capacity
Z
HðT Þ ¼ Hð0Þ þ C p dT ð1Þ

The consequences for protein
stability may seem counterintuitive
but should not be unexpected.
Now, 250 years later, we are still intrigued by this, and the
properties of water continue to present fascinating challenges.
Much of this has been reflected in the discussions surrounding
the heat capacity of proteins and the heat capacity changes
that accompany protein transitions and interactions. In what
follows, I shall try to present a (mostly personal) view of the
current situation in relation to heat capacity effects in protein
unfolding and related processes in solution, mainly from a
physical chemistry perspective. More comprehensive and
authoritative reviews can be found elsewhere.2,3
Z
SðT Þ ¼ Sð0Þ þ ðC p =T Þ dT ð2Þ
where Cp is the heat capacity at constant pressure, T is the
absolute temperature, and the integral is taken from absolute
zero (0 K) to the required temperature. H(0) and S(0) are the
zero-point enthalpy and entropy, respectively. (Equivalent
expressions exist for heat capacity at constant volume, Cv.
Here, however, we shall normally assume constant pressure
because that is what mostly applies. Except for gases, the
numerical difference between Cp and Cv is negligible for most
practical purposes.)
For enthalpy (H=U þ PV, with the internal energy, U, cor-
rected for pressure-volume effects), the physical interpretation
Received Date: August 27, 2010
Accepted Date: November 1, 2010
Published on Web Date: November 04, 2010

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Figure 1. Typical DSC data for the unfolding of a small globular
protein (lysozyme) in solution at various pH values.8 The increase
in area under each endotherm with higher Tm and the higher heat
capacity baselines after the unfolding transitions are both indica-
tions of the significant positive ΔCp commonly associated with
such processes. (Adapted from ref 8, with permission; protein
structure drawn from pdb 1HEW.9)
of eq 1 is straightforward; starting from absolute zero,
each increment in temperature, dT, requires addition of
heat energy, dQ=Cp dT, with summation up to the required
temperature to give the total enthalpy. The entropy relation-
ship, eq 2, is perhaps less intuitive but derives from the Sec-
ond Law definition of the entropy increment, dS = dQ/T =
Cp dT/T. In molecular thermodynamics/statistical mechanics
terms, this relates to the number of ways in which heat
energy may be distributed within the object.
Heat capacity also plays a fundamental role in determining
the size of thermodynamic fluctuations in any system. All
objects are subject to thermal fluctuations arising from
Brownian-motion-like bombardment from surrounding atoms
and molecules. For any object of mass m and specific heat cv
(heat capacity per unit mass), the mean-square internal
energy fluctuations at temperature T are given by
ÆδU 2 æ ¼ kB T 2 mcv
where kB is the Boltzmann constant. For most everyday
objects, these fluctuations are so small (in proportion to the
total energy) as to go unnoticed most of the time. However, for
much smaller objects, particularly microscopic or mesoscopic
systems of the size of protein molecules, these fluctuations are
relatively large.4,5 This is why protein structures must be con-
sidered as dynamic objects at the molecular level.
For any thermodynamic transition (A f B) in which the
different states (A, B) have different heat capacities, it follows
from eqs 1 and 2 that the enthalpy and entropy changes for
the transition will also be temperature-dependent
Z
ΔHðA f BÞ ¼ H B -H A ¼ ΔHð0Þ þ ΔC p dT ð4Þ
Z undergoing reversible unfolding, typically at low pH and low
ΔSðA f BÞ ¼ S B - S A ¼ ΔSð0Þ þ ðΔC p =T Þ dT
where ΔCp =Cp,B - Cp,A is the change in heat capacity of the
system in going from A to B and ΔH(0) and ΔS(0) are the
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Figure 2. Examples of commonly observed heat capacity effects
associated with protein unfolding in solution. (A) DSC data for
thermal denaturation of yeast phosphoglycerate kinase, illustrat-
ing exothermic baseline distortion and heat capacity decrease
caused by irreversible precipitation of unfolded protein. (B) Re-
peat DSC scans of thermal unfolding of lysozyme, showing gradual
accumulation of misfolded forms, thought to arise from cis-trans
isomerization of proline peptide bonds at high temperature.10
Note how the apparent heat capacities for the less-well-folded
forms (below Tm) are higher than those in the original native state.
transition enthalpy and entropy changes at 0 K (or any other
convenient reference temperature).
Equation 4 is often used in its differential form as a con-
ð3Þ venient way to estimate ΔCp from the temperature depen-
dence of observed heats of reaction
ΔC p ¼ DΔH=DT ð6Þ
Protein Heat Capacity ; The Experimental Situation. The
measurement of heat capacities and heat capacity changes
for proteins in dilute (<1 mg mL-1) aqueous solutions is now
relatively straightforward using commercially available differ-
ential scanning calorimeters (DSC) specifically designed for
the purpose, most usually in the 0-110 C temperature
range.6,7 “Typical” DSC data for the thermal unfolding of a
simple globular protein in solution are shown in Figure 1. This
illustrates various features that we shall consider in a moment,
but first, a note of caution. The sort of behavior shown in
Figure 1 is usually only seen for relatively small proteins
ð5Þ concentration. Unfortunately, most proteins do not behave
that way, especially near neutral pH. Unfolded protein is sticky
stuff, and protein denaturation is frequently accompanied by
aggregation or other irreversible effects. This gives rise to
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Figure 3. DSC thermogram of the thermal aggregation of insulin
in solution under conditions where the unfolded protein under-
goes a kinetically limited exothermic nucleation-growth transition
to a condensed protein aggregate similar to amyloid fibrils.12,13 The
inset shows the more typical thermogram seen for protein un-
folding (without aggregation). Note the decreased heat capacity
for the fibril state (arrow). (Adapted from ref 12, with permission;
structures drawn from pdb files 4INS14 and 2KIB.15)
experimental artifacts, as indicated in Figure 2, that can make
accurate heat capacity analysis difficult. However, sufficient
data on reasonably representative samples are now available
so as to make thermodynamic analysis appropriate.
Unfolded protein is sticky stuff.
For well-behaved proteins undergoing reversible thermal
transitions, the heat capacity curves obtained by DSC
(Figure 1) show several characteristic features. Starting from
low temperature, the apparent protein heat capacity gradually
increases until the onset of thermal unfolding, accompanied
by a sharp peak in Cp corresponding to an endothermic un-
folding transition. The peak of the transition gives the mid-
point temperature (Tm), and the area under the transition
gives the enthalpy of unfolding (ΔHunf) at this temperature.
The heat capacity of the protein above the transition, once it
has returned to baseline, is typically higher than would be
anticipated from extrapolation of the low-temperature base-
line. Consequently, the thermal unfolding shows a positive
heat capacity change, ΔCp. This is also indicated by the
marked increase in the area under the transition peak
(ΔHunf) with increasing Tm. In practice, because post-transi-
tion baselines are often poorly defined, it is customary to
perform a series of experiments, varying conditions such as
pH, to give ΔHunf over a range of Tm values, from which ΔCp
may be estimated using eq 6. The validity of this approach
has been demonstrated by Pfeil and Privalov11 and can be
justified because the heats of proton ionization of acidic
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Figure 4. Typical thermodynamic data for reversible two-state
unfolding of a globular protein, with Tm = 40 C, ΔHunf(Tm) =
300 kJ mol-1, and ΔCp = 9 kJ mol-1 K-1.
amino acid side chains is generally small with respect to
ΔHunf.
This positive ΔCp has been taken as a defining feature of
protein unfolding in solution, but it is possibly better to ob-
serve that the lower heat capacity of folded protein (below Tm)
is rather more characteristic of a condensed state of the
polypeptide. This is indicated by DSC data on aggregated
protein (Figures 2 and 3), showing that condensed protein
phases, either nonspecific aggregates (Figure 2) or more
ordered amyloid-like precipitates (Figure 3) as well as the
natively folded state (Figure 1), all show a relatively low heat
capacity in comparison with the more dynamic, unfolded
polypeptide.
As a consequence of the positive ΔCp, the thermody-
namics of the protein unfolding transition (assuming a simple
two-state equilibrium) show significant temperature effects,
as illustrated in Figure 4. Historically, thermodynamic stability
profiles such as these were first obtained by indirect spectro-
scopic analyses, predating more direct calorimetric observa-
tions, and are a tribute to the careful measurements done at
that time.16 What they show is that both enthalpic (ΔHunf) and
entropic (TΔSunf) contributions increase markedly with tem-
perature, but in a compensatory fashion such that the unfold-
ing free energy (ΔGunf = ΔHunf - TΔSunf) is relatively small
and much less sensitive to temperature variation. This “thermo-
dynamic homeostasis”8 may be functionally useful and is
one example of more ubiquitous “enthalpy/entropy com-
pensation” effects now seen in many systems.8,17,18 Here, it
is simply a consequence of a positive ΔCp, which, by virtue of
the fundamental thermodynamic relationships, eqs 4 and 5,
requires that both ΔH and ΔS increase with temperature, with
approximately linear correlation over narrow temperature
ranges.8,18 The midpoint temperature of the unfolding transi-
tion, Tm, equivalent to the “melting point” in conventional
phase transitions, where ΔGunf = 0, is given by the point at
which the ΔHunf and TΔSunf lines intersect. Because the ΔH
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Figure 5. Comparison of experimental specific heats (heat capa-
city, Cp) at 298 K for pure organic solids and liquids24,25 (top panel)
with folded and unfolded proteins in aqueous solution26 (bottom
panel). Experimental data are for compounds up to and including
C5 with elemental composition CHO, CHN, or CHNO, for which
data are available (118 crystalline solids and 159 pure liquids). The
mean values ((s.d.) are 1.32 ((0.22) and 2.18 ((0.38) J K-1 g-1
for solids and liquids, respectively. Mean Cp values for native (n =
35) and denatured (n = 13) proteins are 1.49 ((0.13) and 1.95
((0.15) J K-1 g-1 for native and denatured states, respectively. The
curves show Gaussian fits to the individual distributions. (Adapted
from ref 24, with permission.)
and TΔS lines are almost parallel on this scale (Figure 4), the
intersection point (Tm) will be very sensitive to relatively small
perturbations, analogous to the familiar moir e effect, where
slight changes in displacement between almost parallel lines
gives rise to much larger changes in fringe patterns. This
indicates why it is so difficult to predict the thermal stability of
proteins, and I am not aware of any molecular dynamics
or other theoretical simulations that have yet addressed this
seriously.
The curvature of the free-energy line in the thermal stabil-
ity profiles (Figure 4) is a direct consequence of the finite
positive ΔCp, and as a result, proteins have a temperature of
maximum thermodynamic stability (maximum ΔGunf) for
folding. ΔGunf falls off either side of this maximum, becoming
negative (i.e., thermodynamically favorable for unfolding) not
only at high temperatures, as would be expected, but also, by
extrapolation, at low temperatures. This implies that native
proteins should unfold at sufficiently low temperatures,
though this temperature is usually below the freezing point
of water and experimentally inaccessible under normal con-
ditions. Alternatively, starting with unfolded protein at low
temperature, the protein should fold spontaneously as the
temperature is raised. The demonstration of “cold denaturation”
in certain cases was a satisfying verification of these
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predictions,19 though the structure of the cold denatured
state is ambiguous.20
Anomaly? What Anomaly? The conclusions arising out of
the positive ΔCp for protein unfolding are in many ways
counterintuitive. For example, the positive slope of the ΔHunf
versus T plot (Figure 4) means that unfolding gets more
endothermic with increasing temperature; that is, the higher
the temperature, the more heat energy that is needed to
unfold the protein. Conversely, at sufficiently low tempera-
tures, ΔHunf becomes negative, and exothermic unfolding
(cold denaturation) becomes a possibility. Such apparently
anomalous effects were initially unexpected and led to much
discussion and (sometimes) confusion. However, they were
taken as a striking vindication of the hypothesis that protein
folding was driven by hydrophobic interactions or, more
cautiously, that a positive ΔCp reflects exposure of hydropho-
bic groups upon unfolding. For example, to quote Brandts and
Hunt in 1967, “...More specifically, there is an anomalously
large heat capacity associated with the unfolded protein
which apparently reflects a striking alteration in the process
of accommodation of the exposed hydrophobic side chains in
the solvent phase with changing temperature...”.21 The hydro-
phobic effect22 and its possible role in biomolecular interac-
tions was highlighted by Walter Kauzmann's influential
review,23 laying the foundations for much of what has been
discovered since. However, it is interesting to note that, even
then, Kauzmann was careful to point out that unusual thermo-
dynamic effects are not just restricted solely to hydrophobic
groups, and that similar thermodynamic signatures could be
inferred for interactions involving hydration of salts and other
polar groups; “It is evident that both salt linkages and hydro-
phobic bonds are stabilized predominantly by entropy effects
rather than by energy effects.”23 Therefore, let us re-examine
the situation as regards protein heat capacity.
Unusual thermodynamic effects
are not just restricted solely
to hydrophobic groups.
Comparison with heat capacities of other substances is
informative. Figure 5 shows the absolute specific heats (heat
capacity per gram) of various folded and unfolded proteins in
solution together with a range of organic compounds in the
solid or liquid state at room temperature. As would be
expected from the increase in degrees of freedom in the fluid
state, organic liquids tend to have a higher specific heat
capacity than solids. Data for proteins in solution fall in the
same range, with Cp for folded proteins somewhat higher, on
average, than organic solids and the more flexible, unfolded
proteins slightly lower than organic liquids. One might ratio-
nalize this, at least qualitatively, by simply observing that the
folded protein state is probably a bit more flexible and
dynamic than a crystalline organic solid, whereas the un-
folded protein, though undoubtedly even more flexible and
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Figure 6. Comparison of the specific heat capacity increments
upon melting of simple compounds (right) with protein unfolding
(left).24 Absolute heat capacities for pure solid and liquid com-
pounds are plotted with respect to the normal melting point (ΔT =
T - Tm). For simplicity, the heat capacity plots for simple com-
pounds omit the large spike in Cp associated with heat uptake at
the melting point, ΔT = 0. For proteins, being smaller cooperative
units, this heat uptake takes place over a wider temperature range,
as indicated by the dashed line. (Adapted from ref 24, with
permission.)
dynamic, is still a polymer and will have fewer available
degrees of freedom than a small molecule in the true liquid
phase. (Bear in mind also that thermally unfolded protein still
retains considerable structure which, though dynamic and
heterogeneous, is a long way from true random coil polymer.)
To get a rough idea of the numbers involved, consider the
following. Classical equipartition theory would give an aver-
age thermal energy of order ½kBT for each thermally
accessible degree of freedom, corresponding to a Cp of around
½kB, or ∼4 J mol-1 K-1 per degree of freedom. For a typical
amino acid (mean residue weight ≈ 110 Da), this converts to a
specific heat change of around 0.04 J g-1 K-1 per degree of
freedom. Consequently, the average ΔCp increment of order
0.5 J g-1 K-1 (Figure 5) upon protein unfolding would
translate to an increase of about 12 degrees of freedom per
amino acid residue in this crude estimate. This seems not
unreasonable. Consider also the classic empirical Dulong and
Petit law, also rationalized by thermodynamic equipartition
theory, which gives an upper limit to the heat capacity (for
solids) of 3R (=3NAkB =25 kJ mol-1 K-1) per mole of atoms.
For a typical atomic weight of 14 (e.g., C, N, and O in proteins,
ignoring H), this corresponds to a specific heat of ∼1.8 J g-1 K-1,
which is remarkably (and probably fortuitously) close to
the observed values for organic liquids and unfolded protein
(Figure 5). Heat capacities for real solids (and folded
proteins?) would naturally be lower at normal temperatures
because of quantum limitations on the accessibility of vibra-
tional degrees of freedom. Of course, order-of-magnitude
estimates such as these have ignored complexities arising
from intermolecular interactions in the liquid state and/or
solvation effects, but such effects will generally only serve to
replace one kind of degree of freedom with another, with
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relatively little effect on heat capacity estimates at this crude
level of approximation.
It is therefore interesting to note that, if viewed as a simple
order-disorder transition, the heat capacity increase upon
protein unfolding is entirely unexceptional, with experimen-
tal values lying comfortably within the range that would be
expected from the properties of organic materials in general
(Figure 5), both theoretically and experimentally. This is
further indicated by comparison (Figure 6) with the heat
capacity increments upon melting of a range of simple
compounds, including water. Note that the ΔCp upon melting
of polar substances is significantly greater than that for simple
hydrocarbons, consistent with our understanding of residual
hydrogen-bonded structures in polar liquids, water being the
prime (but not only) example. Again, plotted on the same
scale, the ΔCp for protein unfolding is unexceptional. There-
fore, where is the anomaly? Furthermore, does this mean that
we might reasonably treat protein unfolding as a simple
“melting” process, albeit with quite small lumps of material
immersed in a solvent environment?
The heat capacity increase upon
protein unfolding is entirely
unexceptional.
Of course, protein unfolding is not really a phase transition
in the classic sense. The size of the cooperative unit is finite,
determined by the size of the macromolecule in the simplest
case, and this is why the thermal unfolding transition of the
protein is relatively broad compared to the (infinitely) sharp
cooperative melting transitions of crystalline solids. Moreover,
the protein molecules are in solution, with all of the potential
solvation/hydration issues that this entails. Indeed, that is the
picture that has dominated much of the thinking about
protein folding thermodynamics, driven by burial of hydro-
phobic side chains in the relatively nonpolar interior of the
globular protein structure. However, when I look inside of a
protein structure, it does not look particularly “hydrophobic”,
certainly not to the extent that one sees inside of a detergent
micelle, for example. Yes, there are more nonpolar groups
inside, but maybe not so many as one might expect, and with
every amino acid side chain, polar or otherwise, there comes
the polar amide backbone. In structural terms, this is what
usually dominates our description of the folded protein, an
intricate, closely packed network of hydrogen bonds, mostly
involving backbone -NH and -CdO groups, in which the
overriding rule seems to be “let no H-bond go unsatisfied”.
Potential voids in this structure are filled by side chains,
usually nonpolar (to avoid breaking the rule), but almost as
an afterthought ; a topologically complex, three-dimen-
sional jigsaw puzzle in which the main aim is to fill space as
efficiently as possible to give a densely packed structure.
Maybe this is the clue. Such structures inevitably rely on
cooperativity; not until the last piece is in place does the
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structure stabilize. Partially folded structures just will not hang
together in the face of constant thermal buffeting; it is either
folded or it is not. (Okay, the two-state model for protein
folding is an approximation, and the real picture is one of a
complex, multidimensional conformational space popu-
lated by different distributions, but the simpler view will
serve for now.) Perhaps, in terms of physical chemistry at
least, we should look upon the folded protein molecule as
just another (albeit small) chunk of organic solid. This is not a
new idea (very little is in this Perspective). It has long been
recognized that the molecular packing densities within
globular proteins are close to what is found in organic
solids.27,28 The adiabatic compressibilities and thermal ex-
pansion coefficients of protein molecules are likewise com-
parable to organic solids.29 So why not heat capacity?
Please be aware that I am being deliberately provocative
here. The conventional approach has been to explain ΔCp (and
other thermodynamic properties) in terms of changes in the
environment of polar and nonpolar groups, often taking a two-
dimensional analysis based on changes in solvent-accessible
surface areas (ΔASA). This seems reasonable and has the
advantage that it can be parametrized, at least in principle, by
comparison with the thermodynamics of small-molecule trans-
fer or by statistical analysis of ΔCp data from proteins of known
structure.2,3,30,31 However, this approach attributes essentially
all of the effects to solvation changes, potentially at the expense
of the neglect of more global, three-dimensional contributions
from conformational dynamics and other protein intramolec-
ular interactions.8,12,24 It is possibly for this reason that area-
based models for ΔCp have come to widely differing conclu-
sions about the relative contributions of polar versus nonpolar
groups, for example.2,3 All ΔASA models agree that exposure of
nonpolar groups during protein unfolding should result in an
increase in heat capacity, as classically expected for hydropho-
bic groups, though absolute magnitudes vary. However, totally
disparate results are obtained for polar groups, with no agree-
ment as to the sign, let alone the magnitude of the ΔCp, to be
attributed to their exposure. This probably reflects the paucity of
data for real proteins and the lack of appropriate model systems
in a statistically underdetermined system, together with un-
certainties about ΔASA estimates in the absence of information
about polypeptide conformations in the thermally denatured
state. In the most comprehensive analysis to date, Robertson
and Murphy3 showed that there is little statistically significant
difference between polar and nonpolar groups; both make
similar positive contributions to ΔCp, as might be expected
from fundamental principles.8 The best correlation is with the
overall size or number of amino acid residues in the protein,
regardless of their polarity.3
For simplicity, and in keeping with the theme of this issue,
I have limited discussion to protein (un)folding. However,
similar heat capacity effects are seen in most protein-protein
and protein-ligand interactions, regardless of whether they
involve hydrophobic groups or not.12 The consequences can
be quite extreme, with heats of binding varying significantly
and sometimes even changing sign over relatively small
temperature ranges. This implies that enthalpy (ΔH) mea-
surements at a single temperature might be less informative
than is sometimes assumed.
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Therefore, although the heat capacities of proteins are
perhaps less anomalous than was once thought, they are still
fundamental to our understanding of the thermodynamics of
protein folding and binding. In the absence of suitable empiri-
cal models, one might hope that accurate molecular dynamics
simulations or other computational methods could provide
some guidance. However, this remains a major computational
challenge. Although some progress is being made in estimation
of free-energy changes, no realistic simulation has yet modeled
the experimentally observed heat capacity effects and asso-
ciated variations in enthalpy and entropy. This is understand-
able; adequate computation at just one temperature is hard
enough, especially with inclusion of solvent; therefore, repeti-
tion at different temperatures just increases the burden. Inter-
estingly, however, the direct relationship between heat capacity
and energy fluctuations (eq 3) might provide a convenient and
computationally economical method for estimation of Cp in
properly configured molecular dynamics simulations. The
“noise” in such simulations at a single temperature, once
equilibrated with an appropriate thermal bath, should relate
directly to the heat capacity of the system. This would probably
be most easily implemented in a ligand binding simulation
where the energy fluctuations in the free protein at a particular
temperature should be reduced in the (usually) more rigid
ligand-bound state. Calculation of the resulting decrease in heat
capacity (negative ΔCp), using eq 3, in comparison with that in
experiment, would be an interesting exercise.
AUTHOR INFORMATION
Corresponding Author:
*E-mail: alanc@chem.gla.ac.uk. Phone: (þ44) 141 330 5278. Fax:
(þ44) 141 330 4888.
Biographies
Alan Cooper is Professor of Biophysical Chemistry at the Uni-
versity of Glasgow. Following B.Sc. (Theoretical Physics) and Ph.
D.(Biophysics) studies at Manchester, he did postdoctoral work at
Oxford and Yale, laying foundations for a lifetime's fascination (and
frustration) with the thermodynamics of biomolecular structures
and their interactions. His association with Joseph Black's old
department is entirely serendipitous. By a further quirk of history,
his maternal ancestors were neighbors of Joseph Black, working
nearby in Glasgow as impoverished handloom weavers. Personal
website: http://www.chem.gla.ac.uk/staff/alanc/
ACKNOWLEDGMENT Much of this work evolved under the
auspices of the Biological Microcalorimetry Facility in Glasgow,
funded by the U.K. Biotechnology and Biological Sciences
Research Council, and I am grateful to numerous colleagues and
collaborators over many years, none of whom are to be blamed for
the crazy ideas presented here.
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