Академический Документы
Профессиональный Документы
Культура Документы
Email alerting service Receive free email alerts when new articles cite this article - sign up
in the box at the top right-hand corner of the article or click here
In this paper, we give an overview of our studies by static and time-resolved X-ray
diffraction of inverse cubic phases and phase transitions in lipids. In §1, we briefly discuss
the lyotropic phase behaviour of lipids, focusing attention on non-lamellar structures, and
their geometric/topological relationship to fusion processes in lipid membranes. Possible
pathways for transitions between different cubic phases are also outlined. In §2, we discuss
the effects of hydrostatic pressure on lipid membranes and lipid phase transitions, and
describe how the parameters required to predict the pressure dependence of lipid phase
transition temperatures can be conveniently measured. We review some earlier results of
inverse bicontinuous cubic phases from our laboratory, showing effects such as pressure-
induced formation and swelling. In §3, we describe the technique of pressure-jump
synchrotron X-ray diffraction. We present results that have been obtained from the lipid
system 1 : 2 dilauroylphosphatidylcholine/lauric acid for cubic–inverse hexagonal,
cubic–cubic and lamellar–cubic transitions. The rate of transition was found to increase
with the amplitude of the pressure-jump and with increasing temperature. Evidence for
intermediate structures occurring transiently during the transitions was also obtained. In
§4, we describe an IDL-based ‘AXCESS’ software package being developed in our
laboratory to permit batch processing and analysis of the large X-ray datasets produced
by pressure-jump synchrotron experiments. In §5, we present some recent results on the
fluid lamellar–Pn3m cubic phase transition of the single-chain lipid 1-monoelaidin, which
we have studied both by pressure-jump and temperature-jump X-ray diffraction. Finally,
in §6, we give a few indicators of future directions of this research. We anticipate that the
most useful technical advance will be the development of pressure-jump apparatus on
the microsecond time-scale, which will involve the use of a stack of piezoelectric pressure
actuators. The pressure-jump technique is not restricted to lipid phase transitions, but can
be used to study a wide range of soft matter transitions, ranging from protein unfolding
and DNA unwinding and transitions, to phase transitions in thermotropic liquid crystals,
surfactants and block copolymers.
Keywords: lyotropic liquid crystals; phase transitions; hydrostatic pressure;
X-ray diffraction
(a) (b)
Figure 1. (a) The fluid lamellar La phase. (b) Curvature frustration in a fluid bilayer upon heating.
(a) (b)
Lmax
Lmin
rw
Figure 2. (a) The inverse hexagonal HII phase. (b) Packing frustration in the inverse hexagonal
phase where a is the lattice parameter; rw, the water core radius; and Lmax and Lmin, the maximum
and minimum chain extension, respectively.
elastic energy and the two halves of the bilayer form parallel interfaces to the
underlying minimal surface (Templer et al. 1998a). This degeneracy is broken if
the two halves of the bilayer tend to form constant curvature interfaces instead.
Since the three underlying minimal surfaces pack space differently (degree of
compactness increasing from P–D–G), the degeneracy may also be broken by
transverse interactions between the lipid layers across the water regions. The
phase sequence with decreasing water content is thus expected to follow the
order Im3m–Pn3m–Ia3d (Templer et al. 1998a; Schwarz & Gompper 2000), and
this is observed experimentally (Templer et al. 1998b). The Ia3d (G) cubic phase
is very common both in type I and type II versions in surfactants, lipids and
block copolymers (Bates & Fredrickson 1999), whereas Pn3m (D) and Im3m (P)
seem only to occur as type II (inverse) versions in lipid systems so far. We (and
Figure 3. The underlying minimal surfaces and skeletal graphs (centres of water channels) for the
inverse bicontinuous cubic phases. (a) Pn3m (D), (b) Ia3d (G) and (c) Im3m (P).
others) have found that for various lipid systems which exhibit La–HII transitions
upon heating, reducing the chain length, typically to approximately C12, usually
induces the formation of inverse bicontinuous cubic phases between the La and
the HII phases, or may suppress the HII phase completely.
Many cellular processes including endocytosis, exocytosis and membrane
budding involve changes in membrane topology. The inverse bicontinuous cubic
phases may be relevant to, or may even act as intermediates in some of these
processes, since it is likely that the mechanism of formation of the cubic phase
from a corresponding fluid lamellar phase has much in common with the
mechanism of cell membrane fusion and fission (Siegel & Banschbach 1990).
Much is now known about amphiphile–water mixtures in equilibrium but, as
yet, rather little is known about their non-equilibrium behaviour. This is
surprising given that in all of the examples we have mentioned, the structural
dynamics of phase transitions in the lyotropic mesophases play a profound role.
Previous studies of lyotropic phase transitions have concentrated on
transformations between different lamellar structures and from lamellar to
inverse hexagonal phases, with remarkably little work being done on transitions
involving cubic phases (Erbes et al. 1994; Qiu & Caffrey 2000; Zana 2005).
However, a complete understanding of the physical processes governing such
transitions, including the nature of any intermediates formed and the
mechanistic routes taken, is essential if we are to further our knowledge of
their possible roles in cellular processes involving membranes. Rather little is
known yet about the mechanisms and pathways of transitions involving cubic
phases. It was first thought that they might follow the same pathway as the
Bonnet transformation, a view which was quickly realized to be unphysical,
since it would necessarily involve tearing and self-intersection of the bilayer
(Hyde et al. 1984). Later, it was suggested that a continuous P–D–G transition
could occur by a stretching mechanism, without any tearing (Sadoc &
Charvolin 1989; Benedicto & O’Brien 1997). Thus, a sixfold junction of water
channels in the P (Im3m) phase can be converted into two fourfold junctions of
‘untwist’
Figure 5. Visualization of a possible continuous pathway between the QIIG and QIID cubic phases.
the D (Pn3m) phase; each of these can then, in turn, be converted into two
threefold junctions of the G (Ia3d) phase (figure 4).
Recently, it has been suggested that such continuous cubic transitions could
involve non-cubic (tetragonal, rhombohedral) distortions of the underlying
minimal surfaces, yet with the surfaces remaining minimal during the processes
(Fogden & Hyde 1999; Schroeder et al. 2004). We can try to visualize this
continuous process for the G–D phase transition that involves (figure 5), first, a
tetragonal stretch, changing c/aZ1 to c/aZ2, followed by an ‘untwisting’ of the
water channels with a merging of each pair of three-way junctions to form single
four-way junctions.
(a)
0
−0.002
Cp (cal.°C−1)
−0.004
−0.006
−0.008
(b)
0
−2000
−4000
Q (µcal)
−6000
−8000
−10000
20 30 40 50 60
T (°C)
Figure 6. Calorimetric scans of the gel–fluid transition of DPPC/water. (a) DSC scan; (b) PPC scan.
The tilted gel–rippled gel transition at 358C is detected by DSC but not by PPC.
it is necessary to determine the molar entropy DSm and molar volume DVm
changes at the transitions. These can be conveniently measured using a
combination of differential scanning calorimetry (DSC) for the enthalpy DHm
(figure 6a) and the rather new technique of pressure perturbation calorimetry
(PPC) for the volume change DVm (figure 6b). DSC measures heat flow to/from
the sample upon a change of temperature, whereas PPC measures the heat flow
induced by a small change in pressure, which is directly proportional to the
coefficient of thermal expansion aZ(1/V )(vV/vT)p (Lin et al. 2002). The DSC
values from the scan shown in figure 6a for the transition temperature Tt
and enthalpy change DHm at the gel–fluid (chain-melting) transition of the
lipid dipalmitoylphosphatidylcholine (DPPC) in excess water are 41.58C and
33.8 kJ molK1, respectively, yielding a transition entropy DSm of 108 J KK1 molK1.
The PPC value from the scan shown in figure 6b for the volume change DVm is
23.5 cm3 molK1. In combination, our data predict a pressure dependence of the
gel–fluid transition of DPPC/water of 228C kbarK1, in perfect agreement with
the measured pressure dependence of the transition temperature for this lipid
(Kusube et al. 2005). The Clapeyron equation predicts a linear relationship
between pressure and transition temperature when DSm and DVm are
independent of pressure, or have the same pressure dependence; in practice,
the linearity is often maintained until at least 2 kbar.
There is an interesting consequence of the fact that the Clapeyron equation
contains the ratio of the volume change and the entropy change; a phase
transition with a smaller enthalpy (entropy) change, such as a fluid–fluid
transition, also tends to have a correspondingly smaller volume change, and thus
the pressure dependence of the transition temperature is little changed. In
practice, for fluid lamellar–bicontinuous cubic, lamellar–hexagonal and hexago-
nal–inverse micellar cubic phases, the value of dTt/dp lies within the narrow
range of 20–308C kbarK1 (Duesing et al. 1997).
The possibility that pressure might induce topological transitions in
membranes was suggested by Gruner and co-workers, although for the lipid
system studied, they concluded that pressure increased the rate of formation of
the cubic phase, rather than inducing its formation (So et al. 1993). For the
phospholipid ditetradecylphosphatidylethanolamine/H2O, we found that QIID and
QIIP inverse bicontinuous cubic phases (space groups Pn3m and Im3m) were
induced to form between the fluid lamellar La and HII phases when the pressure
was increased above 700 bar (Duesing et al. 1997). On the other hand, for 1 : 2
phosphatidylcholine/fatty acid mixtures in water, having a range of symmetric
chain lengths, pressure-induced formation of cubic phases was not observed
(Winter et al. 1999).
The application of hydrostatic pressure to lyotropic phases in excess water
tends to increase the lattice parameters of the phases. This effect is caused by the
increase in conformational order of the hydrocarbon chains by pressure, and is
the opposite effect to that of increasing temperature. The effect is generally small
for lamellar phases (less than 2 Å kbarK1), slightly larger for the HII phase, but
can be as much as 80 Å kbarK1 for inverse bicontinuous cubic phases, where the
reduction in chain splay tends to reduce the magnitude of the (negative)
interfacial curvature, thereby swelling the phase if it is in contact with an excess
water phase (Duesing et al. 1997; Winter et al. 1999). This can provide a method
for isothermally tuning the diameter of the water channels of lipid bicontinuous
cubic phases, which could be useful for certain biotechnological applications.
To second order in the principal curvatures, the curvature elastic energy per
unit area of a thin amphiphile film is given (Helfrich 1973) by
gc Z 2kðH KH0 Þ2 C kG K; ð2:2Þ
where H and K are the surface-averaged mean and Gaussian curvatures, H0 is the
spontaneous mean curvature, and k and kG are the mean curvature (bending)
and Gaussian curvature (saddle-splay) moduli, respectively. k is a measure of the
energy required to bend the film where kG is a measure of the energy cost of
changing its Gaussian curvature; from the Gauss–Bonnet theorem, this second
term integrates to a constant value unless a change occurs in topology. A positive
value of kG favours saddle deformations of the film, since these have negative
Gaussian curvature K. For systems which tend to form inverse phases, for the
lipid monolayer, the spontaneous curvature H0 is negative and k is positive;
typical values for monoolein are K0.025 ÅK1 and 1.2!10K20 J (approx. 3 kT),
respectively (Vacklin et al. 2000). On the other hand, kG for a lipid monolayer in
an inverse cubic phase is negative, with typical values of K0.75k (Templer et al.
1998c) or K0.83k (Siegel & Kozlov 2004). For a symmetric lipid bilayer, H0
is zero (by symmetry), kb is twice the value for a single monolayer, but a more
complex form is found for the Gaussian modulus,
kbG Z 2ðkG K4kH0 [Þ; ð2:3Þ
where all the terms on the right-hand side, including the thickness [, refer to the
lipid monolayer. Since the second term on the right-hand side is overall positive
for a lipid that tends to form inverse phases (negative monolayer H0), kbG can be
positive even when kG for a monolayer is negative. This is one of the main driving
forces behind the formation of inverse bicontinuous cubic phases in lipid systems
(Seddon & Templer 1993; Templer et al. 1998a).
Our results (Duesing et al. 1997; Winter et al. 1999) indicate that increasing
pressure tends to reduce the magnitude of the spontaneous curvature H0,
increase the bending rigidity k and increase the rigidity of chain extension. We
surmise from the pressure-induced formation of inverse bicontinuous cubic
phases (Duesing et al. 1997) that pressure also tends to increase kbG , and thereby
stabilizing inverse bicontinuous cubic phases; although this is difficult to predict
from equation (2.3), as the effect of pressure on kG is not known, and the effect of
pressure on k and H0 will tend to cancel out. In contrast, Mariani and co-workers
(Mariani et al. 1996; Pisani et al. 2001, 2003) report similar effects on H0, but
deduce that k decreases with pressure, and that the ratio kG/k is positive and
increases strongly with pressure (from 0.14 to 1 by 1.8 kbar).
We have used the X-ray pressure-jump technique (Mencke et al. 1993; Österberg
et al. 1994; Erbes et al. 1996; Winter et al. 1999; Winter 2002; Winter & Koehling
2004) to investigate the rate and mechanism of lyotropic phase transitions in
monoglyceride/water and fatty acid/phospholipid/water systems, by monitoring
the time evolution of these structural conversions using time-resolved X-ray
diffraction. The use of pressure as a trigger mechanism has several advantages:
(i) the solvent properties are not significantly altered, (ii) pressure propagates
rapidly, indicating that equilibrium is achieved rapidly, and (iii) pressure-jumps
can be both in the pressurization and depressurization directions. The Dortmund
millisecond pressure-jump X-ray cell was used for our experiments (Woenckhaus
et al. 2000). This cell has flat diamond windows that can be used at pressures up
to 8 kbar, and the pressure-jumps occur in 5–7 ms. In order to reduce the
attenuation of the X-ray beam by the diamond windows, it is best to carry out
the experiments at short wavelengths, such as 0.75 Å (17 keV), which gives
typically a transmission of 65%. Beamline ID02 at the ESRF is ideally suited to
these experiments, as will be beamline I22 at the Diamond synchrotron, once this
is operational.
We have used time-resolved X-ray diffraction to monitor transitions, induced
by pressure-jumps, between various inverse lyotropic phases of a 1 : 2
dilauroylphosphatidylcholine/lauric acid (DLPC/LA) system in 50 wt% water
(Squires et al. 2000, 2002, 2005; Seddon et al. 2003). The pressure–temperature
(p–T ) phase diagram (figure 7) of this sample was first assessed by static X-ray
diffraction measurements.
65 H2O
+ HO C O
O
HII O
T (°C)
D O O
G -P
+ O O
O
O
D O
HO C
60 N
Lα
0 0.5 1.0
p (kbar)
Figure 7. The pressure–temperature ( p–T ) phase diagram of 1 : 2 DLPC/LA in 50 wt% water. The
exact position of the G–D phase boundary was found to be somewhat dependent on sample history.
800 500
√2
G (Ia3d) √6 D (Pn3m)
600 400
√3
300
400
200
√8 √22
200 √24
√20 √26 √6 √9
√16 100
√4 √8
√14
0
0
Figure 8. Small-angle X-ray diffraction patterns of the G (Ia3d ) and D (Pn3m) cubic phases. Both
exposure times were 0.1 s. The squared ratios of the reciprocal spacings of the observed peaks are
indicated on the integrated one-dimensional patterns (insets). The lattice parameters are 176.2 and
108.5 Å.
The IDL-based AXCESS software package, developed in our laboratory, allows for
batch processing and analysis of the large X-ray datasets produced by pressure-
jump synchrotron experiments. It has a widget-based front end for user
accessibility, and can process thousands of images sequentially, allowing for a
large reduction in analysis time.
Figure 10a shows a small-angle diffraction pattern from a silver behenate test
sample in a highly ordered lamellar mesophase. The program automatically finds
the centre of the image and integrates over a butterfly-shaped area to produce a
(b)
10000
÷2
1000 ÷3
÷4 ÷6 ÷8 ÷9
100
10
28
t (s–1)
59.1
41.4
29.1
20.4
14.4
10.1
7.10
5.00
3.40 ÷22
2.20 ÷6 ÷8 ÷16
1.40 ÷20 ÷24
÷14 ÷26
0.70 (a)
0.20
–0.20
0 0.01 0.02 0.03 0.04 0.05
s (Å–1)
Figure 9. Time-resolved X-ray diffraction plots of the G–D transition induced at TZ59.58C by a
pressure-jump from 600 to 240 bar (Squires et al. 2005). The intensities are plotted logarithmically
to emphasize the weak peaks. The time increment between successive 0.1 s frames increases
geometrically from bottom to top.
Figure 10. AXCESS software package for batch processing/analysis of X-ray patterns.
We have studied the fluid lamellar to Pn3m (QIID ) inverse bicontinuous cubic
phase transition, effected by means of jumps in temperature and pressure, for
the single chain amphiphile monoelaidin in excess water (Conn et al. 2006).
On increasing temperature or decreasing pressure, the system displays the
phase sequence Lb–La–QIIP –QIID –HII (Czeslik et al. 1995). In particular, the fluid
lamellar to Im3m cubic (QIIP ) transition is interesting, as it is not well defined,
proceeding via a kinetically hindered intermediate, termed X, of unknown
structure and characterized by a broad featureless ring of scatter at low angle in
the diffraction pattern (Caffrey 1987).
(a) (b)
1717
806 QIIP(2) QIIP
÷4 647 ÷4 Q D (2)
÷2 ÷2 Q D (2) ÷2
261 ÷4 ÷3 II
÷2 ÷3 II
220 ÷6
÷4
85 ÷6
QIIP(1) 76
t (s–1)
÷2 D
QII(1)
28 ÷3 ÷2 QD(1)
26 ÷3
II
9 9
X X
2 3
Lα Lα
0.005 0.010 0.015 0.020 0.025 0.005 0.010 0.015 0.020 0.025
s (Å–1) s (Å–1)
Figure 11. Pressure-jumps from (a) 1110 to 240 bar and (b) 1110 to 260 bar at TZ46.78C are
shown. Each original two-dimensional X-ray image has been integrated to produce a one-
dimensional plot of intensity versus scattering vector, s, which is then stacked vertically as a
(nonlinear) function of time.
Figure 11 shows the structural changes following pressure-jumps from (a) 1110
to 240 bar and (b) 1110 to 260 bar, at TZ46.78C. The two jumps, carried out on
the same sample, are remarkably similar and are observed to follow a common
mechanistic route. The disappearance of the fluid lamellar phase is accompanied
by the appearance of a broad, featureless ring of scatter (marked X in figure 11).
After a period of a few seconds, the diffuse scatter resolves itself into a set of
peaks from intermediate inverse bicontinuous cubic phases, QIID and QIIP , which
replace it. These intermediate cubic phases are initially significantly more
hydrated than expected from equilibrium results and dehydrate over time. In
both jumps, a final QIID phase forms at fairly constant lattice parameter.
A major and long-standing problem in the field of lipid phase transition kinetics
is the lack of reproducibility in results. The requirement for intense synchrotron
X-ray sources for adequate time resolution indicates that a transition may be
observed only a few times, yet a number of studies have noted an increase in the
speed of transition with successive jumps. We have overcome the variability in
such dynamical measurements and, for the first time, this has allowed us to record
the dynamical process reproducibly in temperature- and pressure-jump experi-
ments. Figure 12 shows the lattice parameters of all phases present during the
transformation, superimposed for both jumps (a) (filled symbols) and (b) (hollow
symbols). With the exception of the QIIP (2) phase, which forms during jump (a) but
not jump (b), the two transformations are virtually identical. We observe the
transition to be highly reproducible not only in terms of the sequence of phases and
their lattice parameters, but also in the kinetics of the process, i.e. the time at
which each successive phase appears. We believe that the effect is owing to the
homogenization of domain size on repeated thermal cycling and strongly linked to
the transfer of water throughout the sample. The relative amounts of each phase
present throughout the transformation may be approximated by the area under
one of the characteristic diffraction peaks.
(a)
180
(b)
160
55.5
55.0
140 54.5
54.0 Lα
a (Å)
53.5
120
53.0
52.5
100 52.0
51.5
–1.0 –0.5 0 0.5 1.0 1.5 2.0
80 t (s–1)
60 Lα
Figure 12. (a) The change in lattice parameter with time following the pressure-jumps shown in
figure 11 superimposed for both jumps (a, 1110 to 240 bar) (filled symbols) and (b, 1110 to 260 bar)
(hollow symbols). (b) The inset is a close-up view of the behaviour of the lamellar phase in the first
2 s following the jump.
Q
Q
Q
Q
I (Q) (arbitrary units)
t (s–1)
Figure 13. The change in relative intensity (area under one of the characteristic diffraction peaks)
of each phase with time following the pressure-jump shown in figure 11a.
6. Future prospects
1680 ÷2
QIID
÷3
1440
1200
÷2 P
960 QII
t (s–1)
÷4
÷6
720
480
240 X
Lα
0.001 0.002 0.003
s (Å–1)
Figure 14. Temperature jump from 30 to 608C, using a laboratory-based X-ray beamline. Each
two-dimensional X-ray image was integrated to produce a one-dimensional plot of intensity versus
scattering vector, s, which was then stacked vertically as a function of time.
180
160
140 QIIP
120
QIID
a (Å)
100
80
60 Lα
40
0 200 400 600 800 1000 1200 1400 1600 1800
t (s–1)
Figure 15. The change in lattice parameter following the temperature-jump shown in figure 14.
Intensity data are not shown owing to the poor resolution of the in-house X-ray equipment used.
be used to study a wide range of soft matter transitions, ranging from protein
unfolding and DNA unwinding, to phase transitions in liquid crystals,
surfactants and block copolymers.
We anticipate that the most useful technical advance will be the development
of X-ray pressure-jump apparatus on the microsecond time-scale, involving the
use of piezoelectric stacks of pressure actuators. Such a device, for pressure-
jumps of up to G200 bar in 80 ms, has recently been developed for optical studies
of protein solutions (Pearson et al. 2002). Beamline I22 at the new Diamond
synchrotron, with its planned fast detectors with enhanced count-rate and
energy-range capabilities, will be an ideal beamline at which to establish a
UK-based microsecond X-ray pressure-jump facility.
This work has been supported by Platform grant (GR/S77721) and Ph.D. studentships from the
EPSRC. We thank Aurelien Huisman (Socrates student, Imperial College London) for measuring
the DSC and PPC scans shown in figure 6, and Nick Brooks, Chandrashekhar Kulkarni, Sarra
Sebai and Christina Turner for their help with carrying out the synchrotron experiments. We
also thank Prof. R. Winter and Ms J. Kraineva (Dortmund University) and Drs S. Finet and
T. Narayanan (ESRF) for their invaluable contributions.
References
Bates, F. S. & Fredrickson, G. H. 1999 Block copolymers—designer soft materials. Phys. Today 52,
32–38.
Benedicto, A. D. & O’Brien, D. F. 1997 Bicontinuous cubic morphologies in block copolymers
and amphiphile/water systems: mathematical description through the minimal surfaces.
Macromolecules 30, 3395–3402. (doi:10.1021/ma9614353)
Caffrey, M. 1987 Kinetics and mechanism of transitions involving the lamellar, cubic, inverted
hexagonal, and fluid isotropic phases of hydrated monoacylglycerides monitored by time-
resolved X-ray diffraction. Biochemistry 26, 6349–6363. (doi:10.1021/bi00394a008)
Charvolin, J. 1990 Crystals of fluid films. Contemp. Phys. 31, 1–17.
Conn, C. E., Ces, O., Mulet, X., Finet, S., Winter, R., Seddon, J. M. & Templer, R. H. 2006
Dynamics of structural transformations between lamellar and inverse bicontinuous cubic
lyotropic phases. Phys. Rev. Lett. 96, 108102-1–108102-4. (doi:10.1103/PhyRevLett.96.108102)
Czeslik, C., Winter, R., Rapp, G. & Bartels, K. 1995 Temperature- and pressure-dependent phase
behaviour of monoacylglycerides monoolein and monoelaidin. Biophys. J. 68, 1423–1429.
Duesing, P., Seddon, J. M., Templer, R. H. & Mannock, D. A. 1997 Pressure effects on lamellar
and inverse curved phases of fully hydrated dialkyl phosphatidyl–ethanolamines and b-D-
xylopyranosyl-sn-glycerols. Langmuir 13, 2655–2664. (doi:10.1021/la970050q)
Erbes, J., Czeslik, C., Hahn, W., Winter, R., Rappolt, M. & Rapp 1994 On the existence of
bicontinuous cubic phases in dioleoylphosphatidylethanolamine. Ber. Bunsenges. Phys. Chem.
98, 1287–1293.
Erbes, J., Winter, R. & Rapp, G. 1996 Rate of phase transformations between mesophases of the
1 : 2 lecithin fatty acid mixtures DMPC/MA and DPPC/PA—A time-resolved synchrotron
X-ray diffraction study. J. Chem. Soc. Faraday Trans. 100, 1713–1722.
Erbes, J., Gabke, A., Rapp, G. & Winter, R. 2000 Kinetics of phase transformations between
lyotropic lipid mesophases of different topology: a time-resolved synchrotron X-ray diffraction
study using the pressure-jump relaxation technique. Phys. Chem. Chem. Phys. 2, 151–162.
(doi:10.1039/a907613a)
Fogden, A. & Hyde, S. T. 1999 Continuous transformations of cubic minimal surfaces. Eur. Phys.
J. B 7, 91–104. (doi:10.1007/s100510050592)
Helfrich, W. 1973 Elastic properties of lipid bilayers: theory and possible experiments.
Z. Naturforsch. 28C, 693–703.
Hyde, S. T., Andersson, S., Ericsson, B. & Larsson, K. 1984 A cubic structure consisting of a lipid
bilayer forming an infinite periodic minimum surface of the gyroid type in the glycerolmonoo-
leate–water system. Z. Kristallogr. 168, 213–219.
Hyde, S. T., Andersson, S., Larsson, K., Blum, Z., Landh, T., Lidin, S. & Ninham, B. W. 1997 The
language of shape. Amsterdam, The Netherlands: Elsevier.
Kusube, M., Matsuki, H. & Kaneshina, S. 2005 Thermotropic and barotropic phase transitions of
N-methylated dipalmitoylphosphatidylethanolamine bilayers. Biochim. Biophys. Acta 1668,
25–32. (doi:10.1016/j.bbamem.2004.11.002)
Lin, L.-N., Brandts, J. F., Brandts, J. M. & Plotnikov, V. 2002 Determination of the volumetric
properties of proteins and other solutes using pressure perturbation calorimetry. Anal. Biochem.
302, 144–160. (doi:10.1006/abio.2001.5524)
Luzzati, V., Vargas, R., Marianii, P., Gulik, A. & Delacroix, H. 1993 Cubic phases of lipid-
containing systems—elements of a theory and biological connotations. J. Mol. Biol. 229,
540–551. (doi:10.1006/jmbi.1993.1053)
Mariani, P., Paci, B., Boesecke, P., Ferrero, C., Lorenzen, M. & Caciuffo, R. 1996 Effects of
hydrostatic pressure on the monoolein–water system: an estimate of the energy function of the
inverted Ia3d cubic phase. Phys. Rev. E 54, 5840–5843. (doi:10.1103/PhysRevE.54.5840)
Mencke, A. P., Cheng, A. & Caffrey, M. 1993 A simple apparatus for time-resolved x-ray
diffraction biostructure studies using static and oscillating pressures and pressure jumps. Rev.
Sci. Instrumen. 64, 383–389. (doi:10.1063/1.1144261)
Österberg, F., Kriechbaum, M., Polcyn, A., Skita, V., Tate, M. W., So, P. T. C., Gruner, S. M. &
Srramilli, S. 1994 Pressure-induced hydration dynamics of membranes. Phys. Rev. Lett. 72,
2967–2970. (doi:10.1103/PhysRevLett.72.2967)
Pearson, D. S., Holtermann, G., Ellison, P., Cremo, C. & Geeves, M. A. 2002 A novel pressure-
jump apparatus for the microvolume analysis of protein–ligand and protein–protein
interactions: its application to nucleotide binding to skeletal-muscle and smooth-muscle myosin
subfragment-1. Biochem. J. 366, 643–651. (doi:10.1042/BJ20020462)
Pisani, M., Bersntorff, S., Ferrero, C. & Mariani, P. 2001 Pressure induced cubic-to-cubic phase
transition in monoolein hydrated system. J. Phys. Chem. 105, 3109–3119.
Pisani, M., Narayanan, T., Di Gregorio, G. M., Ferrero, C., Finet, S. & Mariani, P. 2003 Compressing
inverse lyotropic systems: structural behavior and energetics of dioleoyl phosphatidyl ethanolamine.
Phys. Rev. E 68, 021924-1–021924-11. (doi:10.1103/PhysRevE.68.021924)
Qiu, H. & Caffrey, M. 2000 The phase diagram of the monoolein/water system: metastability and
equilibrium aspects. Biomaterials 21, 223–234. (doi:10.1016/S0142-9612(99)00126-X)
Sadoc, J.-F. & Charvolin, J. 1989 Infinite periodic minimal-surfaces and their crystallography in
the hyperbolic plane. Acta Crystallogr. A45, 10–20.
Schroeder, G. E., Ramsden, S. J., Fogden, A. & Hyde, S. T. 2004 A rhombohedral family of
minimal surfaces as a pathway between the P and D cubic mesophases. Physica A 339, 137–144.
(doi:10.1016/j.physa.2004.03.056)
Schwarz, U. S. & Gompper, G. 2000 Stability of inverse bicontinuous cubic phases in lipid–water
mixtures. Phys. Rev. Lett. 85, 1472–1475. (doi:10.1103/PhysRevLett.85.1472)
Seddon, J. M. & Templer, R. H. 1993 Cubic phases of self-assembled amphiphilic systems. Phil.
Trans. R. Soc. A 344, 377–401.
Seddon, J. M. & Templer, R. H. 1995 Polymorphism of lipid-water systems. In Handbook of
biological physics (ed. R. Lipowsky & E. Sackmann) Structure and dynamics of membranes, vol.
1, pp. 97–160. Amsterdam, The Netherlands: Elsevier. (series ed. A.J. Hoff )
Seddon, J. M., Squires, A., Ces, O., Templer, R. H., Woenkhaus, J. & Winter, R. 2003 Time-
resolved diffraction studies of inverse cubic phases and phase transitions of lipids. In Self-
assembly (ed. B. H. Robinson), pp. 212–221. Amsterdam, The Netherlands: IOS Press.
Siegel, D. P. & Banschbach, J. L. 1990 Lamellar- inverted cubic phase transition in N-methylated
dioleoylphosphatidylethanolamine. Biochemistry 29, 5975–5981. (doi:10.1021/bi00477a014)
Siegel, D. P. & Kozlov, M. M. 2004 The Gaussian curvature elastic modulus of N-monomethylated
dioleoylphosphatidylethanolamine: relevance to membrane fusion and lipid phase behaviour.
Biophys. J. 87, 366–374. (doi:10.1529/biophysj.104.040782)
So, P. T. C., Gruner, S. M. & Shyamsunder, E. 1993 Pressure-induced phase transitions in
membranes Phys. Rev. Lett. 70, 3455–3458. (doi:10.1103/PhysRevLett.70.3455)
Squires, A., Templer, R. H., Ces, O., Gabke, A., Woenckhaus, J., Seddon, J. M. & Winter, R. 2000
The kinetics of lyotropic phase transitions involving the inverse bicontinuous cubic phases.
Langmuir 16, 3578–3585. (doi:10.1021/la991611b)
Squires, A., Templer, R. H., Seddon, J. M., Woenckhaus, J., Winter, R., Finet, S. & Narayanan, T.
2002 Kinetics and mechanism of the lamellar to gyroid inverse bicontinuous cubic phase
transition. Langmuir 18, 7384–7392. (doi:10.1021/la0259555)
Squires, A. M., Templer, R. H., Seddon, J. M., Woenkhaus, J., Winter, R., Narayanan, T. & Finet, S.
2005 Kinetics and mechanism of the interconversion of inverse bicontinuous cubic mesophases.
Phys. Rev. E 72, 011502-1–011502-16. (doi:10.1103/PhysRevE.72.011502)
Templer, R. H., Seddon, J. M., Duesing, P. M., Winter, R. & Erbes, J. 1998a Modelling the phase
behaviour of the inverse hexagonal and inverse bicontinuous cubic phase in 2 : 1 fatty
acid/phosphatidylcholine mixtures. J. Phys. Chem. B 102, 7262–7271. (doi:10.1021/jp972837v)
Templer, R. H., Seddon, J. M., Warrender, N. A., Syrykh, A., Huang, Z., Winter, R. & Erbes, J.
1998b Inverse bicontinuous cubic phases in fatty acid/phosphatidylcholine mixtures. I. The
effects of chainlength, hydration and temperature. J. Phys. Chem. B 102, 7251–7261. (doi:10.
1021/jp972835a)
Templer, R. H., Khoo, B. J. & Seddon, J. M. 1998c Gaussian curvature modulus of an amphiphilic
monolayer. Langmuir 14, 7427–7434. (doi:10.1021/la980701y)
Vacklin, H., Khoo, B. J., Madan, K. H., Seddon, J. M. & Templer, R. H. 2000 The bending
elasticity of 1-monoolein upon relief of packing stress. Langmuir 16, 4741–4748. (doi:10.1021/
la991408g)
Winter, R. 2002 Synchrotron X-ray and neutron small-angle scattering of lyotropic lipid
mesophases, model biomembranes and proteins in solution at high pressure. Biochim. Biophys.
Acta 1595, 160–184.
Winter, R. & Koehling, R. 2004 Static and time-resolved synchrotron small-angle x-ray scattering
studies of lyotropic lipid mesophases, model biomembranes and proteins in solution. J. Phys.:
Condens. Matter 16, S327–S352. (doi:10.1088/0953-8984/16/5/002)
Winter, R., Erbes, J., Templer, R. H., Seddon, J. M., Syrykh, A., Warrender, N. A. & Rapp, G.
1999 Inverse bicontinuous cubic phases in fatty acid/phosphatidylcholine mixtures. II. The
effects of pressure and lipid composition. Phys. Chem. Chem. Phys. 1, 887–893. (doi:10.1039/
a808950g)
Woenckhaus, R., Koehling, R., Winter, R., Thiyagarajan, P. & Finet, S. 2000 High pressure-jump
apparatus for kinetic studies of protein folding reactions using the small-angle synchrotron x-ray
scattering technique. Rev. Sci. Instrumen. 71, 3895–3899. (doi:10.1063/1.1290508)
Zana, R. (ed.) 2005. Dynamics of surfactant self-assemblies. London, UK: Taylor and Francis.
Discussion
H. F. GLEESON (School of Physics and Astronomy, University of Manchester,
UK ). Do you see flow as part of the p-jump or T-jump transition process? Work
of Julia Yeoman (Oxford) simulating flow in cubic (blue phase) structures show
structural changes in the unit cell—could this kind of process be contributing to
the intermediate signals in the transitions from cubic to cubic?
J. M. SEDDON. In our experiments, we have been careful to create sample
environments, in which no shearing of the hydrated lipid samples can occur,
nor is extraneous water able to flow in or out of the samples. However, in
transforming from one phase to another there may indeed be local flows of
moduli, kG and kbG . The monolayer Gaussian modulus kG has been found to be
negative for systems forming inverse curved phases, and with a magnitude less
than that of the bending modulus k. The bilayer Gaussian curvature modulus kbG
is directly related to the monolayer Gaussian modulus, but also includes a term
that contains the bending modulus k. The explicit expression (Helfrich &
Rennschuh 1990) is
Additional references
Hed, G. & Safran, S. A. 2003 Initiation and dynamics of hemifusion in lipid bilayers. Biophys. J. 85,
381–389.
Helfrich, W. & Rennschuh, H. 1990 Landau theory of the lamellar-to-cubic phase-transition.
J. Phys. 51, 189–195.
Shearman, G. C., Ces, O., Templer, R. H. & Seddon, J. M. 2006 Inverse lyotropic phases of lipids
and membrane curvature. J. Phys.: Condens. Matter. 18, S1105–S1124. (doi:10.1088/0953-
8984/18/28/S01)