RESEARCH COMMUNICATIONS

The maximum energy Wmax an electron may acquire is Evidence for bound and free water
given by
species in the hydration shell of an
Wmax = mc 2 ∆γ , ≈ 2eE0cγ r2 /ω , (29) aqueous micelle
where γ r2 = (1 − β 2 ) −1 . If the length required by an elec-
tron to acquire Wmax amount of energy is LA, then we can Sundaram Balasubramanian†,*, Subrata Pal# and
write Wmax = eE0 L A , which gives LA = 2cγ 0 /ω p . Biman Bagchi#,*
†
The amplitude of the Langmuir wave, driven by beat- Chemistry and Physics of Materials Unit, Jawaharlal Nehru Centre for
Advanced Scientific Research, Jakkur, Bangalore 560 064, India
ing a mm wave with a wiggler, scales as one-third power #
Solid State and Structural Chemistry Unit, Indian Institute of Science,
of wiggler strength and one-sixth power of mm wave Bangalore 560 012, India
power density. It is sensitive to cyclotron resonance.
The electric field amplitude of the Langmuir wave is
Atomistic molecular dynamics simulations revealed
|E | = kφF, where F = |φ /φ0 |. For a typical case of 36 GHz,
the existence of bound and free water molecules in the
mm wave with power density P = 1 MW/cm2, wiggler hydration layer of an aqueous micelle. The bound
strength Bw = 1kG, the value of Langmuir wave electric water molecules can be either singly or doubly hydro-
field amplitude turns out to be 30 MV/m at ωL/ωC ~ 0.8. gen bonded to the polar head group on the surface of
The breaking of the Langmuir wave does not occur here, the micelle. The ratio of bound to free water is found
as the phase velocity of the wave is close to c; wave to be approximately equal to 9 : 1 at 300 K.
breaking occurs when the electron excursion equals a
wavelength. The field amplitude could be enhanced near WATER, inevitably present at the surface of biological
cyclotron resonance, but as one approaches the cyclotron macromolecules and self-organized assemblies, plays a
resonance, the phase velocity of the Langmuir wave critical role in the structure, stability and function of
decreases unless one increases kw. Alternatively, large these systems1–2. However, the layer of water that sur-
phase velocities of the Langmuir wave can be achieved rounds these systems is rather thin, typically 1–3 layers
by propagating the Langmuir wave through a plasma of thick. Thus, the study of hydration layer has turned out to
slightly higher density after it has been generated in the be rather difficult. Dielectric relaxation studies measure
wiggler region. The large amplitude Langmuir wave the collective response of the whole system and, there-
could be utilized for electron acceleration. The length of fore, are not a sensitive probe of the dynamics of the
the plasma column required for maximum particle acce- hydration water. NMR techniques (NOE and NMRD)
leration scales linearly with the wavelength of the mm have the required spatial resolution, but lack the dynamic
wave. The requirement of a large guide field is a serious resolution. Neutron-scattering techniques are beginning
drawback of the scheme. to be applied to study the dynamics of hydration layer in
these systems3. Solvation dynamics may be a reasonably
1. Tajima, T. and Dawson, J. M., Phys. Rev. Lett., 1979, 43, 267.
good probe because it can have both the temporal and
2. Katsouleas, T. and Dawson, J. M., ibid, 1983, 51, 392. spatial resolution4–6; yet it provides only a collective res-
3. Bobin, J. L., Proceedings of ECFA-CAS/CERN-In-2P3-IRF/ ponse. Under these circumstances, computer simulation
CEA – EPS Workshop (ed. Turnet, S.), Orsay, Italy, 1987, p. 58. can play an important role in understanding the nature of
4. Nakajima, K., Phys. Rev. Lett., 1992, 45, 1149; Nakajima, K. the hydration water.
et al., ibid, 1995, 74, 4428.
5. Kitagawa, Y. et al., ibid, 1992, 68, 48.
As the hydration layer is spatially heterogeneous even
6. Rosenbluth, M. N. and Liu, C. S., ibid, 1972, 29, 701. on a molecular-length scale and because the microscopic
7. Clayton, C. E., Joshi, C., Darrow, C. and Umstadetr, D., ibid, interactions are quite complex, a purely analytical study
1985, 54, 2343. of this system is prohibitively difficult. In order to cap-
8. Amiranoff, F. et al., ibid, 1995, 74, 5220. ture some of the basic physics of the dynamics in such
9. Rosenweig, J., Murokh, A. and Pellegrini, C., ibid, 1995, 74,
2467.
systems, a model in terms of a dynamic exchange bet-
10. Kimura, W. D. et al., ibid, 1995, 74, 546. ween bound and free water molecules7 has been pro-
11. Yoder, R. B., Marshall, T. C. and Hirshfield, J. L., ibid, 2001, 86, posed. The bound water molecules are those which are
1756. singly or doubly hydrogen bonded to the protein or to the
12. Liu, C. S. and Tripathi, V. K., Interaction of Electromagnetic surface of a self-assembled aggregate, while the free
Waves with Electron Beams and Plasmas, World Scientific,
Singapore, 1994.
water molecules are not. The microscopic dynamical
event is an exchange between these two states of water
ACKNOWLEDGEMENTS. This work is supported by DST, Govt of within the hydration layer. The model predicts the emer-
India. gence of a slow decay when the binding energy is high.
Received 11 March 2002; revised accepted 2 December 2002

*For correspondence. (e-mail: [email protected]; [email protected].

ernet.in)

428 CURRENT SCIENCE, VOL. 84, NO. 3, 10 FEBRUARY 2003

 RESEARCH COMMUNICATIONS

In this limit, the time constant of the slow decay is just a

the inverse of the rate of bound to free transition7.
Although this model has been semi-quantitatively succes-
sful in explaining a wide range of behaviour, the basic
assumption of the existence of bound and free water
molecules remained unsubstantiated.
In addition to the above, bound and free water mole-
cules give a convenient way to categorize the water mole-
cules in the hydration layer. The bound molecules can be
further sub-divided into two categories – singly hydrogen
bonded or doubly hydrogen bonded. We shall denote
these two by IBW1 and IBW2 respectively. Free water
molecules are denoted by IFW (interfacial free water).
Recently, we have presented several studies aimed at
b
understanding various aspects of interfacial water8. These
were based on detailed atomistic molecular dynamics
(MD) simulations of an anionic micelle, CsPFO (cesium
pentafluorooctanoate). These studies have confirmed the
existence of slow water and ion dynamics in the inter-
facial region. We have also studied the lifetime of hydro-
gen bonds that the water molecules form with the (a)
micellar polar head groups and found that it becomes
considerably longer than those between two water mole-
cules in the bulk.
In the present work, we have extended our previous
study to investigate in detail, the equilibrium structure of
the interfacial water. Our study has clearly revealed that
the water at the interface of CsPFO consists of three dif-
ferent species – IBW2, IBW1 and IFW in the ratio 1.1 : Figure 1. Schematic description of the environment around bound
8 : 0.9, i.e. the bound to free water ratio is 9 : 1. This interfacial waters; a, IBW1, and b, IBW2. Numerical values of the
large ratio is a signature of the highly polar character of geometrical parameters are average values obtained from the MD run.
Water molecules and surfactant head groups are rigid entities in the
the anionic micelle. interaction model. PHGO denotes the oxygen atom of the polar head
As details of the simulation have been discussed else- group of the surfactant, and PHGC denotes the carbon atom in the head
where8, we directly proceed to discussion of the results. group. WO and WH denote the oxygen and hydrogen atoms of the
interfacial water respectively. Broken lines between PHGO and WH
Figure 1 illustrates the average geometries adopted by denote hydrogen bond.
IBW1 and IBW2 water species. Figure 1 gives all the
details about average bond lengths and bond angles cal-
culated from the simulation. There are features which are
absent in IBW1 but present in IBW2, such as a well-
defined distance between the water and the second (non-
bonded) oxygen of the polar head group. This feature at
4.7 Å is prominent for IBW2 water molecules, but is
absent for IBW1. An examination of the full distribution
of bond lengths and bond angles, and not just their aver-
ages, shows that the environment around bound water
species is, in general, more well-defined relative to a
water molecule in pure water.
Figure 2 presents the monomer energy distribution for
all the three species, as also for water molecules in the
bulk. It is seen that the interfacial water molecules have
peaks at lower energies – the IBW2 species has the low-
est potential energy. It is also worth noting that the bound
water molecules have considerably lower energy values
than the free water molecules. It is this enhanced stability
which makes the bound species identifiable, even when Figure 2. Distribution of monomer energies of interfacial water mole-
they are transient because of the dynamic exchange bet- cules compared to that of bulk water.

CURRENT SCIENCE, VOL. 84, NO. 3, 10 FEBRUARY 2003 429

RESEARCH COMMUNICATIONS
27, 203–208; Marchi, M., Sterpone, F. and Ceccarelli, M., J. Am.
Chem. Soc., 2002, 124, 6787–6791.
3. Ruffle, S. V., Michalarias, I., Li, J. and Ford, R. C., J. Am. Chem.
Soc., 2002, 124, 565–569.
4. Sarkar, N., Dutta, A., Das, S. and Bhattacharyya, K., J. Phys.
Chem., 1996, 100, 15483–15486.
5. Jordanides, X. J., Lang, M. J., Song, X. and Fleming, G. R., J. Phys.
Chem. B, 1999, 103, 7995–8005.
6. Riter, R. E., Willard, D. M. and Levinger, N. E., ibid, 1998, 102,
2705–2714.
7. Nandi, N. and Bagchi, B., ibid, 1997, 101, 10954–10961.
8. Balasubramanian, S. and Bagchi, B., ibid, 2001, 105, 12529–12533;
Balasubramanian, S. and Bagchi, B., ibid, 2002, 106, 3668–3672;
Pal, S., Balasubramanian, S. and Bagchi, B., J. Chem. Phys., 2002,
117, 2852–2859; Balasubramanian, S., Pal, S. and Bagchi, B., Curr.
Sci., 2002, 82, 845–854; Balasubramanian, S., Pal, S. and Bagchi,
Figure 3. Schematic description of free energy (solid line) and inter- B., Phys. Rev. Lett., 2002, 89, 115505-1–115505-4.
nal energy (dashed line) profiles of interfacial water species. The spe-
cies are in dynamical equilibrium with themselves and with water
present in the bulk region of the micellar solution. The reaction coordi- Received 24 October 2002; revised accepted 13 January 2003
nate is arbitrary and does not imply any distance. Barrier heights are
also arbitrary. ACKNOWLEDGEMENT. This work is supported by grants from the
Department of Science and Technology and the Council of Scientific
and Industrial Research, to both S.B. and B.B.
ween the free and bound species. A large part of this sta-
bilization comes from the stronger hydrogen bonds that
bound water molecules form with the surfactant head
groups.
Figure 3 provides a schematic of the free energies of Effect of sodium butyrate on
the three species, calculated from their average concen- methylation pattern of retinoblastoma
trations. Despite the reduced monomer energy arising out
of two water–head group hydrogen bonds, the IBW2 state (RB1) gene in human colon tumour cell
is less stable than the IBW1 state due to entropic consi- line HT29
derations (less number of suitable configurations). The
reversible reactions between these states of water on such Mohan L. Gope# and Rajalakshmi Gope#,†,*
a surface should determine the dynamical response of #
Creighton University School of Medicine, Creighton Cancer Center,
interfacial water. Omaha, NE 68178, USA
†
Note that Figure 3 describes the free energy and not the Present address: Department of Human Genetics, National Institute of
Mental Health and Neuro Sciences, Bangalore 560 029, India
binding energy of the three species. The binding energy can
be inferred from Figure 2. The total binding energy of the
IBW2 species is indeed larger than that of IBW1. Sodium butyrate is known to induce morphological
In conclusion, we note that the existence of identifiable and biochemical changes associated with differentia-
bound and free water molecules on the surface can indeed tion in some colon tumour cell lines, including HT29.
help in developing a phenomenological description of In this study the HT29 colon carcinoma cells were
dynamics of water at complex interfaces. The 9 : 1 ratio induced to differentiate with sodium butyrate treat-
ment. High molecular-weight DNA from the untreated
obtained (for IBW and IFW) is expected to be typical for
and sodium butyrate-treated HT29 cells were isolated
ionic micelles. However, this ratio is bound to decrease at different time periods. The DNA was digested with
substantially for proteins due to the existence of hydro- the restriction enzymes MspI and HpaII to reveal
phobic and less polar amino acid groups on its surface. changes in the methylation pattern at the RB1 gene
The water on the protein/membrane surface is expected locus. Sodium butyrate-treated HT29 cells showed an
to play a critical role in the molecular recognition of increase in the level of methylation of retinoblastoma
hydrophobic patches by incoming ligands or drug mole- (RB1) gene compared to the untreated control. The
cules. Work in this direction is under progress. results suggest that the development of human colonic
neoplasia may result from hypomethylation of RB1
1. Nandi, N., Bhattacharyya, K. and Bagchi, B., Chem. Rev., 2000, gene.
100, 2013–2045.
2. Ringe, D., Curr. Opin. Struct. Biol., 1995, 5, 825–829; Sansom, CYTOSINE methylation in CpG dinucleotides is an impor-
M. S. P., Srivastava, I. H., Ranatunga, K. M. and Smith, G. R., tant control mechanism of the transcriptional regulation
Trends Biochem. Sci., 2000, 25, 368–374; Teeter, M. M., Yamano,
in eukaryotes. The transcriptional activity of several
A., Stec, B. and Mohanty, U., Proc. Natl. Acad. Sci. USA, 2001, 98,
11242–11247; Tarek, M. and Tobias, D. J., Phys. Rev. Lett., 2002,
88, 138101-1–138101-4; Mattos, C., Trends Biochem. Sci., 2002, *For correspondence. (e-mail: [email protected])

430 CURRENT SCIENCE, VOL. 84, NO. 3, 10 FEBRUARY 2003