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stability
Susanne Moelbert
Institut de theorie des phenom`enes physiques, Ecole polytechnique federale de Lausanne, CH-1015 Lausanne, Switzerland
B. Normand
Departement de Physique, Universite de Fribourg, CH-1700 Fribourg, Switzerland
arXiv:cond-mat/0305204v2 [cond-mat.soft] 25 Jun 2004
Eob ,qob
A. Hydrophobic-Polar Model
Eos ,qos
The driving force in the process of solvation and ag-
gregation is the effective hydrophobic interaction between
polar water and the non-polar solute [23, 30]. As noted in FIG. 3: Energy levels of a water site in the bimodal MLG
Sec. I, the origin of this interaction is the rearrangement model. The energy levels of shell water are different from bulk
of water around the solute particle. For solute surfaces of water due to breaking and rearrangement of hydrogen bonds
sufficiently high curvature to permit (partial) cage forma- in the proximity of a solute particle. The solid arrows rep-
tion, this process decreases the enthalpy due to reinforced resent the effect of a kosmotropic cosolvent in creating more
hydrogen bonds between water molecules in the solvation ordered states, and the dashed arrows the opposite effect of
shell of the hydrophobic particle in comparison to those chaotropic species.
in bulk water, but reduces simultaneously the number of
degenerate states of the solvent. These physical features
are described by the model of Muller, Lee and Graziano shown that this framework yields a successful description
(MLG), where the energy levels and respective degenera- of preferential binding, and of the resulting destabilisa-
cies of water molecules are determined by the local water tion of aggregates of solute particles as a consequence of
structure [37, 38]. the role of chaotropic cosolvents in reducing the forma-
Because solute particles are relatively large compared tion of water structure. We begin with a brief review of
with single water molecules, we use an adapted version of the model to summarise its physical basis and to explain
the MLG model in which each site contains a group of wa- the modifications required for the inclusion of cosolvent
ter molecules. The distinction between solvent molecules effects.
and non-polar solute particles lies in their ability to form The microscopic origin of the energy and degener-
hydrogen bonds. The continuous range of interaction en- acy parameters for the four different types of water
ergies within a partially hydrogen-bonded water cluster site (ordered/disordered and shell/bulk) is discussed in
may be simplified to two discrete states of predominantly Refs. [36, 41, 42], in conjunction with experimental
intact or broken hydrogen bonds [39]. The water sites in and theoretical justification for the considerations in-
the coarse-grained model may then be characterised by volved. Here we provide only a qualitative explanation
two states, where an ordered site represents a water of their relative sizes, which ensure the competition of
cluster with mostly intact hydrogen bonds, while a dis- enthalpic and entropic contributions underlying the fact
ordered site contains relatively fewer intact hydrogen that the model describes a closed-loop aggregation re-
bonds (Fig. 1). The use of a bimodal distribution in the gion bounded by upper and lower critical temperatures
adapted model is not an approximation, but a natural and densities [40]. Considering first the energies of wa-
consequence of the two-state nature of the pure MLG ter sites (groups of molecules), the strongest hydrogen
model [36]. In the presence of a non-polar solute a fur- bonding arises not in ordered bulk water, but in the
ther distinction is required, between bulk water sites shell sites of hydrophobic particles due to the cage for-
undisturbed by the solute particles and shell sites in mation described in Sec. I (which can be considered as
their vicinity whose hydrogen bonding is altered. As ex- a result of the orientational effect of a hydrophobic sur-
plained in more detail below, this adaptation of the MLG face). Both types of ordered water cluster are enthalpi-
model contains the essential features required to encap- cally much more favourable than disordered groups of
sulate the enthalpy/entropy balance which is the basis molecules, for which the presence of a non-polar parti-
for the primary phenomena of hydrophobic aggregation cle serves only to reduce hydrogen bonding still further,
in two-component water/solute mixtures. The model has making the disordered shell energy the highest of all. As
been used to reproduce the appearance of upper (UCST) regards the site degeneracies, which account for the en-
and lower (LCST) critical solution temperatures and a tropy of the system, cage formation permits very few
closed-loop coexistence regime [40], demonstrating that different molecular configurations, and has a low degen-
the origin of the hydrophobic interaction lies in the al- eracy. Water molecules in ordered clusters have signif-
teration of water structure. icantly reduced rotational degrees of freedom compared
The adapted MLG model has been extended to provide to disordered ones, with the result that the degeneracy of
a minimal model for the study of chaotropic phenomena the latter is considerably higher. Finally, the degeneracy
in ternary water/solute/cosolvent systems [36]. It was of a disordered shell site is still higher than that of a bulk
4
site because the relative reduction of hydrogen bonding for the (partial) formation of hydrogen-bonded cages be-
due to the non-polar surface admits a higher amount of come inappropriate for larger curvatures, on the order
rotational freedom. of 1 nm, where a crossover occurs to depletion or
Figure 3 summarises the energy levels of a water site, dangling-bond models in which hydrogen-bond forma-
which are arranged in the sequence Eds > Edb > Eob > tion is frustrated [50]. While the adapted MLG model be-
Eos . Their respective degeneracies are qds > qdb > qob > comes inapplicable for such particles at the point where a
qos , where the states are denoted ds = disordered shell, clathrate description breaks down, we note [46] that most
db = disordered bulk, ob = ordered bulk and os = or- solute species, and in particular proteins, are atomically
dered shell (cage conformation). We stress that these rough, in that they composed of chains and side-groups
sequences are neither an assuption of the model nor may with high local curvatures on the order of 0.3-0.5 nm.
they be altered if the model is to represent a system As a consequence the model may remain valid for solute
exhibiting the closed-loop form of the coexistence curve particles of considerably greater total dimension. For
for aggregation: only these specific sequences reproduce the purposes of the present analysis, we note that a sig-
appropriately the competition of enthalpic and entropic nificant quantity of the available data concerning cosol-
contributions to the free energy. From the microscopic vent effects has been obtained for protein solutions, and
considerations of the previous paragraph these sequences that the majority of this data is consistent with our ba-
are entirely plausible, and they have been confirmed by sic picture of the dominance of water structure effects in
a range of experimental measurements; the exact values dictating solute solubility. However, while most proteins
of the parameters are not important for the qualitative may indeed fall in the class of systems well described by a
properties of the system while the order of energies and clathrate picture, we stress that the adapted MLG model
degeneracies is maintained. We return in Sec. V to the cannot be expected to provide a complete description for
issue of departure from these sequences. large solute molecules containing many amino-acids of
In the calculations to follow we have used the en- different local hydrophobicity and chemical interactions.
ergy values Eds = 1.8, Edb = 1.0, Eob = 1.0 and
Eos = 2.0, which are thought to be suitably represen-
tative for aqueous solutions, and which have been suc- B. Cosolvent Addition
cessful in describing a number of different types of so-
lution [40, 41, 43]. Their corresponding degeneracies, In comparison with the hydrophobic solute particles,
normalised to a non-degenerate ordered shell conforma- cosolvent molecules are generally small and polar, and
tion, are taken as qds = 49, qdb = 40, qob = 10 and are therefore included directly in water sites by changing
qos = 1 [41]. We remind the reader that these energy and the number of states of these sites. A site containing wa-
degeneracy parameters are independent of temperature ter molecules and a cosolvent particle is referred to as a
and solute density. Their relative values have been found cosolvent site. Kosmotropic cosolvents, being more polar
to be appropriate for reproducing the primary qualitative than water, increase the number of intact hydrogen bonds
features of hydrophobic interactions [40, 41], swelling of at a site [19]. In the bimodal MLG framework their addi-
biopolymers [43], protein denaturation [44, 45], and mi- tion to weakly hydrogen-bonded, disordered clusters may
celle formation [46]. The essential behaviour obtained be considered to create ordered clusters with additional
within this model is indeed found to be rather insensitive intact hydrogen bonds. The creation of ordered states
to the precise parameter values, which may, however, be from disordered ones in the presence of a kosmotropic
refined by comparison with experimental measurements cosolvent increases the degeneracy of the former at the
to yield semi-quantitative agreement for different solu- expense of the latter. This feature is incorporated by
tions [40, 41, 43]. The energy scale is correlated directly raising the number of possible ordered states compared to
with a temperature scale, which we define as kB T 1 . the number of disordered states (solid arrows in Fig. 3),
We conclude this subsection by qualifying the range of
validity of the adapted MLG model, including what is qob,k = qob + b , qdb,k = qdb b ,
meant above by small solute particles. The model is qos,k = qos + s , qds,k = qds s , (1)
not appropriate for solute species smaller than a group
of water molecules with intercluster hydrogen-bonding, where k denotes the states of water clusters containing
which necessitates a linear cluster (and solute) size of kosmotropic cosolvent molecules, and the total number
at least 2 water molecules. The model will also fail to of states is kept constant.
capture the physics of large, uniform systems where the The effect of chaotropic cosolvents on the state de-
linear cluster size exceeds perhaps 10 water molecules, generacies is opposite to that of kosmotropic substances,
because in this case the majority of the water molecules and may be represented by inverting the signs in eq. 1.
in a shell site would correspond effectively to bulk Chaotropic cosolvents are less strongly polar than wa-
water. Most molecular-scale treatments of water struc- ter, acting in an aqueous solution of hydrophobic parti-
ture in the vicinity of non-polar solute particles assume cles to reduce the extent of hydrogen bonding between
a spherical and atomically flat solute surface. These water molecules in both shell and bulk sites [23, 51].
analyses [47, 48, 49] indicate that clathrate models Within the adapted MLG framework, this effect is re-
5
produced by the creation of disordered states, with addi- where a = k, c denotes sites containing water and a kos-
tional broken hydrogen bonds and higher enthalpy, from motropic or chaotropic cosolvent molecule. The site vari-
the more strongly bonded ordered clusters (dashed ar- able i is defined
Q as the product of the nearest neigh-
rows in Fig. 3), whence bours, i = hi,ji n2j , and takes the value 1 if site i is
completely surrounded by water and cosolvent or 0 oth-
qob,c = qob b , qdb,c = qdb + b ,
erwise. The first sum defines the energy of pure water
qos,c = qos s , qds,c = qds + s . (2) sites and the second the energy of cosolvent sites. Be-
We stress three important qualitative points. First, cause a water site i may be in one of q different states,
the cosolvent affects only the number of intact hydrogen i,os is 1 if site i is occupied by water in one of the qos
bonds, but not their strength [23], as a result of which, ordered shell states and 0 otherwise, and i,ds is 1 if it is
also in the bimodal distribution of the coarse-grained occupied by pure water in one of the qds disordered shell
model, the energies of the states remain unchanged. Co- states and 0 otherwise. Analogous considerations apply
solvent effects are reproduced only by the changes they for the bulk states and for the states of cosolvent sites.
cause in the relative numbers of each type of site [eqs. (1) A detailed description of the mathematical structure of
and (2)]. Secondly, cosolvent effects on the relative de- the model is presented in Refs. [40] and [36].
generacy parameters are significantly stronger in the bulk The canonical partition function of the ternary N -site
than in the shell, because for obvious geometrical reasons system, obtained from the sum over the state configura-
related to the orientation of water molecules around a tions {i }, is
non-polar solute particle [36, 41], many more hydrogen-
P Q ni (ni +1)
(1i )
ni (ni +1)
i
bonded configurations are possible in the bulk. Finally, ZN = Zs 2
Zb 2
{ni } i
as suggested in Sec. I, our general framework does not ni (ni 1) ni (ni 1)
(1i ) i
include the possibility of cosolvent-solute interactions, Zs,a 2
Zb,a 2
, (4)
which have been found to be important in certain ternary
systems, specifically those containing urea [52, 53]. where Z = qo eEo + qd eEd for the shell ( =
The illustrative calculations in Secs. III and IV are per- s) and bulk ( = b) states both of pure-water and of
formed with b = 9.0 and s = 0.1 in eqs. (1) and (2). In cosolvent sites ( = s, a and = b, a).
the analysis of Ref. [36] these values were found to pro- When the number of particles may vary, a chemical
vide a good account of the qualitative physics of urea as a potential is associated with the energy of particle addi-
chaotropic cosolvent; in Sec. V we will return to a more tion or removal. The grand canonical partition function
quantitative discussion of the role of these parameters. of the system for variable particle number becomes
By using the same values for a hypothetical kosmotropic X X gc
cosolvent we will demonstrate that a symmetry of model = eNw +(+)Na ZN = eHeff [{ni }] . (5)
degeneracy parameters does not extend to a quantitative, N {ni }
or even qualitative, symmetry in all of the relevant phys-
ical phenomena caused by structure-changing cosolvents. Nw denotes the number of pure water sites, Na the num-
We comment briefly that the fractional value of s arises ber of cosolvent sites and Np the number of solute par-
only from the normalisation convention qos = 1. ticle sites, the total number being N = Nw + Na + Np .
The variable represents the chemical potential associ-
ated with the addition of a water site to the system and
C. Mathematical Formulation the chemical potential for the insertion of a cosolvent
molecule at a water site. From the role of the two cosol-
To formulate a description of the ternary solution vent types in enhancing or disrupting water structure, at
within the framework of statistical mechanics, we begin constant solute particle density is expected to be neg-
by associating with each of the N sites of the system, ative for kosmotropic agents and positive for chaotropic
labelled i, a variable ni , which takes the values ni = 1 ones.
if the site contains either pure water, ni = 0 if the site
contains a hydrophobic particle, or ni = 1 if the site
contains a water cluster including one or more cosolvent D. Monte Carlo Simulations
molecules. This system is described by the Hamiltonian
We describe only briefly the methods by which the
X
N
ni (ni +1)
H[{ni }, {i }] = [(Eob i,ob +Edb i,db )i model may be analyzed; full technical details may be
i=1
2 found in Ref. [40]. Because our primary interest is in
the local solute and cosolvent density variations which
+(Eos i,os + Eds i,ds )(1 i )]
demonstrate aggregation and preferential phenomena, we
X
N
ni (ni 1) focus in particular on Monte Carlo simulations at the
+ [(Eob i,ob,a + Edb i,db,a )i level of individual solute molecules. Classical Monte
i=1
2
Carlo simulations allow the efficient calculation of ther-
+(Eos i,os,a + Eds i,ds,a )(1 i )], (3) mal averages in many-particle systems with statistical
6
FIG. 5: Closed-loop coexistence curves obtained by Monte FIG. 6: -T phase diagram of hydrophobic solute in an aque-
Carlo simulations for an aqueous solution of hydrophobic par- ous cosolvent mixture for two different kosmotropic cosol-
ticles with the same parameters as in Fig. 4. vent concentrations k , obtained by Monte Carlo simulations.
The endpoints of the finite transition lines correspond to the
UCST and LCST.
gregates and a lower value 1 , which may be interpreted
as a strengthening of the hydrophobic interactions gen-
erated between solute particles. of finite length terminated by the UCST and LCST de-
However, the mean-field approximation neglects fluc- marcate the aggregation phase transition. An increase in
tuations in the density, and consequently is unable to cosolvent density leads to a larger separation of UCST
detect intrinsically local phenomena such as preferential and LCST as a direct result of the expansion of the ag-
exclusion of cosolvent particles from the solvation shell of gregation regime (Fig. 5). The transition line is shifted
a hydrophobic particle (Fig. 2). Because the kosmotropic to higher values of , indicating an increased resistance
effect is strongly dependent upon local enhancement of of the system to addition of water at constant volume
water structure, a full analysis is required of the spatial in the presence of cosolvent. This is a consequence of
density fluctuations in the system. We thus turn to the the fact that the resistance to addition of hydrophobic
results of Monte Carlo simulations, which at the macro- particles depends on both and : with increasing co-
scopic level show the same enhancement of hydrophobic solvent concentration, decreases (i.e. becomes more
aggregation (Fig. 5) as indicated in the mean-field anal- negative) and hence must increase for the critical par-
ysis. Below the LCST and above the UCST, one homo- ticle density to remain constant.
geneous mixture is found where the hydrophobic parti- By inspection of the energy levels, and of the alteration
cles are dissolved, while between these temperatures they in their relative degeneracies caused by a kosmotropic
form aggregates of a given density. The LCST and the cosolvent (Fig. 3), it is energetically favourable to max-
critical aggregate densities determined by the mean-field imise the cosolvent concentration in bulk water with re-
calculation do not differ strongly from the numerical re- spect to shell water. Thus one expects a preferential
sults, but the UCST lies at a temperature higher than exclusion, also known as preferential hydration, as repre-
that determined by Monte Carlo simulations. This is to sented schematically in the right panel of Fig. 2. Except
be expected because mean-field calculations neglect fluc- at the lowest temperatures, an increased number of water
tuation effects, generally overestimating both transition molecules in the solvation shell of non-polar solute par-
temperatures. The agreement of the mean-field result for ticles increases the repulsion between solute and solvent,
small k with the simulated value for the LCST suggests resulting in a reduction of solubility, in an enhanced ef-
a predominance of local effects at low temperatures, and fective attraction between solute particles due to the de-
that density fluctuations between sites are small. We creased interface with water as they approach each other,
note, however, that a cosolvent concentration k = 0.61 and thus in aggregate stabilisation.
is required in the mean-field approximation to produce Monte Carlo simulations confirm the expectation that
an expansion of the aggregated phase similar to that ob- the cosolvent concentration is lower in shell water than
served in the Monte Carlo results for k = 0.25, demon- in bulk water. Figure 7 shows the cosolvent shell con-
strating the growing importance of fluctuation effects at centration compared to the overall cosolvent concentra-
higher cosolvent concentrations. tion in the solution, where preferential exclusion of the
The -T phase diagram of the ternary system obtained kosmotropic cosolvent from the solvation shell of the hy-
by Monte Carlo simulations is presented in Fig. 6. Lines drophobic solute particles is observed. At constant chem-
8
2.5
c =0.02
c =0.3
2
c =0.4
c =0.5
c =0.53
T 1.5
0.5
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1
p
3
2
c =0.02
2.5 c =0.3 c, total = 0.03
c =0.4 1.8
2 c =0.5
c, total = 0.28
c,shell /c,total
1.6
1.5 low particle density c, total = 0.35
T
c, total = 0.71
0
0 1 2 3 4 5 6 7 1
0 0.5 1 1.5 2 2.5
T
FIG. 9: -T phase diagram of hydrophobic solute in an aque-
FIG. 10: Preferential binding effect, illustrated by the relative
ous cosolvent mixture for different chaotropic cosolvent con-
concentration of chaotropic cosolvent in the solvation shell of
centrations c , obtained by Monte Carlo simulation. The fi-
hydrophobic particles, for different cosolvent concentrations
nite transition lines terminate at an UCST and a LCST, which
at fixed = 3.0, obtained by pair approximation.
approach each other with increasing cosolvent concentration.
tem in the presence of cosolvent. However, in this case tween the opposing effects of kosmotropic and chaotropic
the endpoints of the lines, which represent the UCST and substances on hydrophobic aggregation of a given solute
LCST for each cosolvent concentration, become closer as species. Indeed, the competition of the two effects has
c increases until they approach a singularity (Fig. 8), been studied for solutions of the protein HLL with the
which we discuss in detail in Sec. V. (denaturing) chaotrope guanidine hydrochloride and the
Figure 10 shows as a function of temperature the co- (native-structure stabilising) kosmotrope betaine [8]. We
solvent concentration in the shell of a solute particle nor- remark, however, that the same cosolvent may have a
malised by the total c . This demonstration of prefer- different effect for solutions of different types and sizes
ential binding of chaotropic agents to non-polar solute of solute (Sec. II), and for this reason a global charac-
particles was obtained from the model of eq. (3) within terisation of the effectiveness of any one agent may be
a pair approximation, which was shown in Ref. [36] to misleading.
provide a semi-quantitative reproduction of the Monte At low cosolvent concentration, the stabilising or
Carlo results. As in Fig. 7, the sharp step at the phase destabilising effects of the two cosolvent types on ag-
transition is due to the sudden change in particle density. gregate formation are rather small, and appear almost
At low particle densities (below the transition) the effect symmetrical in the expansion or contraction of the coex-
is clear, whereas at high densities it is small because the istence region (Figs. 5 and 8, Figs. 6 and 9). For weakly
majority of solvent and cosolvent sites have become shell kosmotropic and chaotropic agents, by which is meant
sites. For low cosolvent concentrations, preferential bind- those causing only small enhancement or suppression of
ing can lead to a strong increase in c on the shell sites aggregation even at high concentrations, such qualitative
(80% in Fig. 10), but the relative enhancement becomes symmetry might be expected as a general feature of the
less significant with increasing total cosolvent concentra- coexistence curves. With the same caveats concerning
tion. Finally, in counterpoint to the discussion at the low concentrations or weak cosolvent activity, this state-
end of Sec. III, we comment that for chaotropic agents, ment is also true for preferential exclusion and binding of
which in bulk water transform ordered states into disor- cosolvent to the solute particles, as shown by comparing
dered ones, preferential binding (cosolvent exclusion) is Figs. 7 and 10. However, differing phenomena emerge for
favoured and solute exclusion decreases. Thus a regime strongly kosmotropic and chaotropic agents.
exists in the p -T phase diagram where at fixed temper-
ature and particle density, aggregates of solute particles
may be made to dissolve simply by the addition of cosol- A. Urea as a Strong Chaotrope
vent.
Chaotropic cosolvents act to suppress the formation of
water structure and, because the latter is already dis-
V. STRONG AND WEAK COSOLVENTS rupted by the presence of a non-polar solute particle, are
more efficient for bulk than for shell sites. Within the
As indicated in Sec. II and illustrated by the results adapted MLG model, a maximally chaotropic agent is
of Secs. III and IV, there is a qualitative symmetry be- one which causes sufficient disruption that the number
10
3
Such singular behaviour is well known in many aque-
(a) ous solutions containing urea, and indeed frequent use
2.5 c = 0.8
is made of the destabilising properties of this cosolvent
c = 0.75 (Sec. I). We stress at this point that the chaotropic ac-
2
c = 0.65 tion of urea is by no means universal: in a number of
T c = 0.5 systems, such as the ultra-small solute methane [20, 54],
1.5
T * c = 0.33 and proteins with variable hydrophobicity [49] or signifi-
c = 0.03 cant interactions between chain segments and the cosol-
1
vent molecules [52, 53], its effect may in fact be inverted.
However, because of its ubiquity as a solubilising agent
0.5
for most binary systems, we continue to consider urea as
0
a generic example of a strong chaotrope.
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 The aggregation singularity was reproduced in the
p* p
qualitative analysis of urea as a chaotropic agent per-
formed in Ref. [36], and is illustrated in Fig. 11. Because
of the discrete nature of the Monte Carlo simulations, the
3 exact parameters for the critical point cannot be found
c =0.03 (b) using this technique; these were instead obtained only
2.5 c =0.33 within the mean-field approximation, and as a result
c =0.5 cannot be considered to be quantitatively accurate for
2 c =0.65 the parameters of the system under consideration. Fig-
c =0.75 p<p*
T c =0.8
ure 11(a) shows the shrinking of the coexistence curves
1.5 with increasing cosolvent concentration until they vanish
c =0.81
at a critical value c = 0.81 (cf. c 0.55 expected from
1 inspection of the Monte Carlo results in Fig. 8). The
p>p*
corresponding evolution of the transition line in the -T
0.5 phase diagram is shown in Fig. 11(b).
With the canonical choice of degeneracy parameters
0
0 2 4 6 8 (Sec. IIB) qds = 49, qdb = 40, qob = 10 and qos = 1 [41],
the values b = 9.0 and s = 0.1 required in eq. (2)
to reproduce this critical vanishing are indeed such that
FIG. 11: (a) Coexistence curves for ternary water/cosol- qob,c = 1 and qos,c = 0.9 become very close. The level
vent/solute systems with different cosolvent concentrations of fine-tuning of the underlying parameters required in
c , obtained by mean-field calculation. At a critical cosolvent the adapted MLG model reflects not a weakness of the
concentration of c = 0.81, the LCST and UCST coincide, at
framework but rather of the strongly chaotropic proper-
a critical temperature T = 1.35 and critical particle density
p = 0.14, where the aggregation regime is reduced to a single ties of urea in water: the destruction of hydrogen-bonded
point. For still higher cosolvent concentrations the solution networks is almost complete at high concentrations. In
is homogeneous over the full range of temperature and par- practice, concentrated urea is used to destabilise aque-
ticle density. (b) Mean-field -T phase diagram for different ous solutions of many folded proteins, micelles and ag-
cosolvent concentrations. As c increases, the separation of gregates of hydrophobic particles. While we are unaware
LCST and UCST decreases until for c = c they meet at the of systematic experimental studies of the effects of urea
temperature TL = TU = T , where the value L = U = 7.05 on the extent of the coexistence regime, aggregate desta-
determines p = 0.14. bilisation has been achieved at high concentrations in
certain systems. At room temperature, the solubility of
the highly hydrophobic amino-acid phenylalanin is dou-
bled in an 8M urea solution (a solution with equal volume
of ordered bulk states is reduced to its hypothetical min-
fractions of urea and water) [16].
imum value, which is equal to the number of ordered shell
states; by the definition based on hydrogen-bond forma-
tion, it is not possible to have fewer ordered bulk than or-
dered shell states, so qob qos . When qob and qos become B. Sodium Chloride as a Weak Kosmotrope
similar, the entropic gain of creating ordered bulk states
by aggregation (removal of shell sites) is largely excluded, Kosmotropic cosolvents increase the extent of water
and thus a strongly chaotropic agent raises the solubility structure formation, raising the number of ordered bulk
of hydrophobic particles very significantly. With a suffi- and shell sites. While this process may be more efficient
cient concentration of such cosolvents, aggregation may for ordered shell states, their degeneracy cannot exceed
be completely prevented at all temperatures and den- that of ordered bulk states. A maximally kosmotropic
sities, and the coexistence regime vanishes at a critical agent is one which causes sufficient structural enhance-
value of cosolvent concentration. ment that the number of ordered bulk states rises to the
11
bimodal model of Muller, Lee and Graziano. This model tween solute particles, thus stabilising their aggregates;
considers two populations of water molecules with differ- similarly, the preferential binding of chaotropic agents
ing physical properties, and generates indirectly the ef- to solute particles reduces the hydrophobic interaction,
fective hydrophobic interaction between solute particles. causing the particles to remain in solution. The essen-
Such a description, based on the microscopic details of tial cosolvent phenomena may thus be explained purely
water structure formation around a solute particle, is ex- by the propensity of these agents to promote or suppress
pected to be valid for small solute species, characterised water structure formation, and the effectiveness of a co-
by lengthscales below 1 nm, and for those larger solute solvent is contained in a transparent way in the micro-
particles whose solubility is dominated by the atomically scopic degeneracy parameters of the solvent states.
rough (highly curved) nature of their surfaces.
In the adapted MLG framework, one may include Our focus on water structure is intended to extract one
the ability of kosmotropic cosolvents to enhance and of of the primary factors determining aggregation and co-
chaotropic cosolvents to disrupt the structure of liquid solvent activity, and is not meant to imply that other in-
water simply by altering the state degeneracies [eqs. (1) teractions are not important. The range of solute species
and (2)] associated with the two populations characteris- and additional contributions which are beyond the scope
ing this structure. As a consequence only of these changes of our analysis includes extremely small solutes, large,
in the properties of the solvent, the model reproduces all atomically flat solute surfaces, solutes with differing sur-
of the physical consequences of cosolvents in solution: face hydrophobicity (such as polar side-groups) and sur-
for kosmotropes the stabilisation of aggregates, expan- faces undergoing specific chemical (or other) interactions
sion of the coexistence regime and preferential cosolvent with the cosolvent. However, from the qualitative trends,
exclusion from the hydration shell of hydrophobic solute and even certain quantitative details, observed in a wide
particles; for chaotropes the destabilisation of aggregates, variety of aqueous systems it appears that the adapted
a contraction of the coexistence regime and preferential MLG model is capable of capturing the essential phe-
binding to solute particles. nomenology of cosolvent activity on aggregation and sol-
The microscopic origin of these preferential effects lies ubility for solutes as diverse as short hydrocarbons and
in the energetically favourable enhancement of hydrogen- large proteins.
bonded water structure in the presence of a kosmotropic Acknowledgments
cosolvent, and conversely the avoidance of its disruption
in the presence of a chaotrope. In a more macroscopic in-
terpretation, the preferential hydration of solute particles We thank the Swiss National Science Foundation for
by kosmotropic agents can be considered to strengthen financial support through grants FNRS 21-61397.00 and
the solvent-induced, effective hydrophobic interaction be- 2000-67886.02.
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