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INTRODUCTION
Lipid rafts are membrane-ordered microdomains which exhibit
a dierent composition and ordering than the surrounding
environment and can phase separate and oat in an otherwise
disordered membrane. These domains carry out a number of
important physiological functions: they host signal transduction
pathways and inuence synaptic transmission, apoptosis,
organization of the cytoskeleton, cell adhesion and migration,
and TCR activation and are involved in protein and lipid
sorting.1,2
Predominant components of lipid rafts are glycolipids as well
as sphingomyelin, cholesterol, and transmembrane proteins.
Such lipid rafts are known to be resistant toward detergents
such as Triton X-100, which might be explained by the ability
of cholesterol to cause a closer packing of neighboring acyl
chains due to its rigid sterol group.3,4 Depending on the sugar
headgroup, glycosphingolipids can be divided in two groups,
ceramides and gangliosides. The lipids are referred to as
gangliosides if the headgroup contains a sialic acid, a negatively
charged sugar group. Glycolipids missing the sialic acid are
called ceramides.5 Glycolipid headgroups are covalently
attached to either a glycerol or a sphingosine backbone,
forming a glycophospho- or glycosphingolipid, respectively. In
glycosphingolipids, a long chain of mostly saturated fatty acids
is linked to the sphingosine backbone over an amide bond
while the sugar headgroup is linked to the hydroxyl group of
the backbone. This sugar headgroup can range from a single
sugar residue to complex oligosaccharide chains.6
It was observed that pure glycolipid membranes exhibit
dierent phase behavior than pure phospholipid membranes.
2015 American Chemical Society
DOI: 10.1021/acs.langmuir.5b01617
Langmuir 2015, 31, 93799387
Langmuir
Article
SIMULATION METHODS
Coarse-grained simulations were employed to study the phase
transition of mixed DPPC/glycolipid membranes and the
inuence of glycolipids on the membrane structure.
Coarse-Grained Simulations. The framework of the
Martini parametrization2729 was used for the coarse-grained
representation of lipids (Figure 1) and surrounding water. In
contrast to the atomistic approach, the coarse-grained Martini
force eld models on average four heavy atoms and the
associated hydrogens by a single interaction center, speeding up
simulations by about 2 orders of magnitude. However, at this
reduced resolution, isomers such as glucose and galactose can
not be distinguished. Here, we used the polarizable version of
the Martini force eld,30 which includes charged particles on a
spring for the water beads to include polarizability. Initial tests
with the standard water model led to a reversed eect for the
addition of charged GM1 lipids on the phospholipid phase
transition as compared to experiment. Additionally, GM1
9380
DOI: 10.1021/acs.langmuir.5b01617
Langmuir 2015, 31, 93799387
Langmuir
Article
SZ =
3
1
cos2 Z
2
2
(1)
describes the angle between the vector along the acyl chain
and the membrane normal. The order parameter was calculated
separately for every 20 ns over the whole trajectory.
DOI: 10.1021/acs.langmuir.5b01617
Langmuir 2015, 31, 93799387
Langmuir
Article
Figure 2. Area per lipid in nm2 for all lipids of the systems of pure
DPPC (top), DPPC + 17% GCER (middle), and DPPC + 17% GM1
(bottom) using polarizable water as a function of temperature in K.
Both cooling (black lines) and heating simulations (blue lines) were
performed at a heating rate of 0.2 K/ns. The transition temperature
was determined as described in the Methods section using a Heaviside
function at the transition point (t functions as red solid lines).
Transition temperatures Tcm and Thm are marked by gray dashed and
solid lines, respectively, and those of the mixed DPPC/glycolipid
systems, by black lines.
Tcm(K)
error (K)
Thm(K)
error (K)
6
6
6
264.5
272.1
278.1
1.2
1.1
0.5
305.0
310.9
323.1
0.2
1.0
1.7
DOI: 10.1021/acs.langmuir.5b01617
Langmuir 2015, 31, 93799387
Langmuir
Article
Figure 5. Average cluster size and the number of clusters during a 600
ns equilibration at 340 K, shown for systems DPPC + 17% GM1 (A),
DPPC + 17% GCER (B), and for comparison for randomly selected
DPPC molecules (17%) in a simulation of a pure DPPC bilayer (C).
Glycolipids/lipids within a distance of 0.65 nm were considered to be
in one cluster.
Figure 4. Density distribution of the dierent membrane components analyzed for the systems DPPC, DPPC + 17% GM1, and DPPC + 17% GCER
for the gel (left) and the liquid-crystalline (right) phase.
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DOI: 10.1021/acs.langmuir.5b01617
Langmuir 2015, 31, 93799387
Langmuir
Article
SAS (nm2)
gel
liquid-crystalline
gel
liquid-crystalline
gel
liquid-crystalline
whole bilayer
upper leaet
lower leaet
gel
liquid-crystalline
DPPC (PW)
0.95
0.38
47.38 (0.28)
72.97 (1.27)
0.93
0.38
47.75 (0.22)
68.11 (1.20)
147.28 (1.53)
137.31 (2.93)
35.12 (16.4)
11.42 (5.51)
18.02 (8.26)
515 (9.3)
892 (26.7)
0.84
0.4
46.23 (0.23)
68.08 (1.16)
151.31 (1.92)
139.00 (3.04)
6.93 (3.21)
3.15 (0.78)
3.25 (1.01)
390 (7.4)
779 (25.1)
434 (8.4)
885 (26.0)
Structural observables were determined for both the gel and the liquid-crystalline phase (initial 100 ns and last 100 ns of the simulation,
respectively). Order parameter SZ describes the average SZ of both hydrocarbon chains for the DPPC lipids in all three systems. The average tilt angle
refers to the headgroups of GM1 and GCER (Methods section). The average cluster size was calculated on the last 100 ns of 600-ns-long
equilibrations at 340 K (SAS = solvent assessible surface; standard deviation given in parentheses).
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DOI: 10.1021/acs.langmuir.5b01617
Langmuir 2015, 31, 93799387
Langmuir
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Figure 7. Number of glycolipidglycolipid, glycolipidDPPC, and glycolipidwater contacts for systems DPPC/GM1 (a) and DPPC/GCER (b)
for heating simulations (PW = polarizable water).
Figure 8. Solvent-accessible surface (SAS) area for DPPC, DPPC + 17% GM1, and DPPC + 17% GCER analyzed for heating simulations (a). The
area per residue for the gel (b) and the liquid-crystalline (c) phase is also shown. Residues 141168 and 309336 represent GM1 (cyan) or GCER
(red) molecules, respectively.
Notes
ACKNOWLEDGMENTS
This work was supported by the German Science Foundation
(DFG) within the Research Training Group 1962 Dynamic
Interactions at Biological Membranes. M.P. was supported by a
scholarship within the Programme to Promote Equal
Opportunities for Women in Research and Teaching (FFL)
of the Friedrich-Alexander University of Erlangen-Nurnberg.
We thank the Marrink Group for sharing the glycolipid Martini
parameters. Computer time was provided by the Computing
Center of the Friedrich-Alexander University of ErlangenNurnberg (RRZE).
REFERENCES
AUTHOR INFORMATION
Corresponding Author
DOI: 10.1021/acs.langmuir.5b01617
Langmuir 2015, 31, 93799387
Langmuir
Article
DOI: 10.1021/acs.langmuir.5b01617
Langmuir 2015, 31, 93799387
Langmuir
Article
9387
DOI: 10.1021/acs.langmuir.5b01617
Langmuir 2015, 31, 93799387