pubs.acs.

org/JPCL Letter

Restructuring of Membrane Water and Phospholipids in Direct

Interaction of Neurotransmitters with Model Membranes Associated
with Synaptic Signaling: Interface-Selective Vibrational Sum
Frequency Generation Study
Biswajit Biswas and Prashant Chandra Singh*
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sı Supporting Information

ABSTRACT: Comprehensive molecular-level understanding of

the role of interfacial water and phospholipids associated with
synaptic membranes during their direct interaction with neuro-
transmitters is essential because of their involvement in synaptic
signaling. Herein, the interfacial regions of the synaptic membranes
mimicking anionic and zwitterionic phospholipids are probed in
the presence of dopamine and serotonin neurotransmitters using
surface-specific vibrational sum frequency generation technique.
Neurotransmitters intrude into the headgroup region of both
zwitterionic and anionic lipids by restructuring the interfacial water
associated with the phospholipids, although the restructuring
mechanism is different for both lipids. Neurotransmitters also
decrease the overall ordering of both the phospholipids probably by creating gauche defects. Neurotransmitters restructure the
surface water, conformation, and the ordering of the hydrocarbon chains of the zwitterionic and anionic phospholipids associated
with synaptic membranes, which could be potentially an important step for synaptic signaling.

N eurotransmitters are an important class of chemicals

which transmit brain signals from one neuron across the
synapse to a target cell.1−5 In the presence of an appropriate
electric field, neurotransmitters burst out of the presynaptic
membrane and communicate with receptors present in the
postsynaptic membrane. It has been acknowledged that
phospholipids associated with the synaptic membrane play a
crucial role in synaptic neurotransmission.6−9 However, the
role of phospholipids in synaptic transmission is mainly studied
in the context of their cooperation with membrane proteins,
and the interaction of neurotransmitters with phospholipids is
assumed to be passive in this process.
Apart from the well-documented lipid−protein interactions,
a significant number of recent studies suggest that specific
neurotransmitter−lipid interactions can also affect the synaptic
signaling process.10−12 Thermochemical and NMR studies of
dopamine neurotransmitter with anionic lipid vesicle suggest
that dopamine binds with the anionic membrane mainly via
electrostatic interaction.13−15 Similarly, serotonin, another
important neurotransmitter, has been found to interact with
the zwitterionic lipid vesicles mainly via hydrogen bonds.16
The fluorescence, calorimetry, neutronscattering and diffrac-
tion studies of melatonin suggest that it gets partitioned into
the membrane, thereby affecting its signaling process.11,17,18 It
has also been suggested that the binding of neurotransmitters
depends strongly on the phospholipid composition of the
membrane and that disbalance in the composition of lipids
causes neurological diseases.10,11,14
MD simulation studies have pointed out that lipophilic
neurotransmitters such as serotonin and dopamine intrude into
the interface of synaptic membranes.10,11,16,19 The interfacial
region of the synaptic membranes contains phospholipids as
well as associated interfacial water. Neurotransmitters have
complex interactions with phospholipids as well as interfacial
membrane water which may affect its functionality.11 Indeed, it
has been already known that the interfacial water beneath the
phospholipid plays a key role in the stability of the membrane
as well as fibrillation and conformation change of different
proteins at the membrane.20−22 Hence, understanding the
structure of phospholipid and its associated interfacial water
during the direct interaction of neurotransmitters with the
membrane is essential to get insight into the synaptic signaling
process. However, earlier experimental studies of neuro-
transmitter binding with lipid could not provide insight into
Received: January 18, 2021
Accepted: March 12, 2021

© XXXX American Chemical Society https://dx.doi.org/10.1021/acs.jpclett.1c00173

2871 J. Phys. Chem. Lett. 2021, 12, 2871−2879
The Journal of Physical Chemistry Letters
the property and structure of the synaptic membrane region
because of the lack of interface selectivity of the techni-
ques.13,14,16
Vibrational sum frequency generation (VSFG) is an
interface-specific and sensitive technique that has been utilized
extensively to decode the properties of several different
complex interfaces.23−32 The VSFG technique has been
applied to understand the structure of surface water at the
surfactant/water, lipid/water, and protein/lipid/water inter-
faces.33−36 Apart from that, the conformational changes of
proteins, as well as structural changes of lipid at different soft
interfaces, has also been investigated using the VSFG
technique.24,33,34,36 Indeed, recently, our group has utilized
the VSFG technique to show that the adsorption of
monomeric α-synuclein reorganizes membrane water depend-
ing on the headgroup of the lipid, which affects the fibrillation
process of protein as well.22
In order to get insight into the structure of phospholipid and
associated surface water during the direct interaction of
neurotransmitters with the membrane, the VSFG spectra of
the anionic (1,2-dipalmitoyl-sn-glycero-3-phospho-rac-1-glyc-
erol sodium salt (DPPG)) and zwitterionic (1,2-dipalmitoyl-
sn-glycero-3-phosphocholine (DPPC)) phospholipid mono-
layer/water interfaces in the absence and presence of
dopamine, as well as serotonin neurotransmitters in the
aqueous phase, have been measured (structures of all the
chemicals are provided in Figure 1). Zwitterionic and anionic
Figure 1. Structure of lipids (anionic and zwitterionic), dopamine,
and serotonin neurotransmitters.
phospholipids are chosen as model membrane systems as they
are the significant contributors of the neuronal synaptic
membranes.10,15,37 Dopamine and serotonin are two important
chemical neurotransmitters whose imbalance causes several
serious diseases, such as Parkinson’s disease, hyperactive and
bipolar disorders, and schizophrenia in humans.9,38 Dopamine
and serotonin exist in cationic form around the physiological
pH as shown in Figure 1.39 The details of the interactions of
neurotransmitters with the headgroup and hydrocarbon chain
of phospholipids as well as interfacial water have also been
supplemented with MD simulation data.
This study suggests that neurotransmitters intrude into the
headgroup of the anionic phospholipid as the NH3+ group of
neurotransmitters screens the negative charge of phospholipid
and decreases the probe length of the interfacial membrane
water. Neurotransmitters also penetrate the headgroup region
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of the zwitterionic phospholipid by the restructuring of surface
water and interact selectively with the negatively charged
phosphate group of net neutral zwitterionic phospholipid. The
interaction of neurotransmitters affects the packing of the
hydrocarbon chain of the anionic and zwitterionic lipids as the
ordering of the hydrocarbon chain of both lipids decreases.
Figure 2a depicts the SSP (S, SFG signal; S, Visible; P, IR)
polarized VSFG spectra of anionic air/DPPG/water interface
without and with different concentrations of dopamine. The
spectral feature in the range of 2800−3000 cm−1 represents the
C−H vibration of the hydrocarbon group of DPPG as
dopamine does not show any VSFG signal in the C−H
stretching region at the air/water interface (Figure S1a),
whereas the spectral feature from 3000 to 3600 cm−1
corresponds to the OH stretch of surface water at the air/
DPPG/water interface. In agreement with earlier studies, the
intensity of the OH stretch of water at the air/DPPG/water
interface gets enhanced ∼10 times compared to the air/water
interface (Figure S1b).34,40,41 Increase in the intensity of the
water signal at the air/DPPG/water interface is due to the
increased probed length of oriented water present in the
electric double layer (EDL) between the negatively charged
headgroup of DPPG and diffused counterions.34,41,42 It is
noteworthy that the water signal in the EDL at the charged
lipid interface has the contribution of χ(3) (third-order
susceptibility) along with the χ(2) signal.43 The intensity of
the OH stretch of water at the air/DPPG/water interface
increases upon the addition of ∼100−200 μM of dopamine
than the case of only air/DPPG/water interface. A similar
observation of the increase in the VSFG signal of water on the
addition of an approximately micromolar concentration of
counterions at charged interfaces has also been observed
earlier, and it has been assigned to the removal of destructive
interference from deeper-lying oriented water because of the
phase-matching condition.43,44 Hence, the increase in the
VSFG signal of water on the addition of micromolar dopamine
at the negatively charged interface can be due to the
interference effect from the deeper-lying oriented water,
although the dopamine has OH groups which can also form
a hydrogen bond with water molecules and may orient them
further to enhance the signal.
In contrast, the intensity of the VSFG signal of water
decreases monotonically with the further addition of dopamine
(800 μM to 50 mM) and almost becomes similar to that of the
air/water interface. On addition of higher concentration of
dopamine, the NH3+ group of dopamine gets attracted to the
negative charge of DPPG by decreasing the probe length of
water molecules present in the EDL, causing a decrease in the
water VSFG signal. At sufficiently high concentration (∼50
mM), the NH3+ group of dopamine almost gets into the
vicinity of the phosphate (PO4−) headgroup of DPPG and
neutralizes its surface charge because the intensity of the VSFG
signal in the hydrogen-bonded region of water (3000−3600
cm−1) at the air/DPPG/water surface with dopamine (∼50
mM) is almost like that in the air/water interface. In contrast
to this study, it has been shown earlier that rather than the
NH3+ group, the guanidinium group of the arginine interacts
preferentially with the negatively charged lipid, which may be
possible because of the enhanced hydrogen bonding capability
of the guanidinium group in comparison to that of the NH3+
group.45
The VSFG spectrum of the C−H stretch of the hydrocarbon
chain of DPPG at the air/DPPG/water interface depicts
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Figure 2. (a) SSP polarized VSFG spectra of the anionic air/DPPG/water interface in the presence of different concentrations of dopamine. The
change in the CH3-ss, CH2-ss, and CH3-ss/CH2-ss of the hydrocarbon chain of DPPG for the various concentrations of dopamine is depicted in
panels b, c, and d, respectively. The surface pressure of DPPG during the measurement was 35 ± 5 mN/m (∼57.2 ± 1.8 Å2/molecule) depicting
the liquid compressed (LC) phase of lipid. The addition of dopamine does not affect the surface pressure of lipid within the uncertainty of the
measurement.
signals at 2850 and 2875 cm−1 along with a relatively broad
peak with double-peak features centered at 2950 and 2965
cm−1, respectively. The spectral features at 2850 and 2875
cm−1 have been attributed to CH2-ss and CH3-ss of the
hydrocarbon chain of the DPPG lipid, whereas the broad
feature containing the peaks around 2950 and 2965 cm−1 have
been assigned to its CH3−FR and CH3-as, respectively.40 In
the presence of dopamine, the intensity of the CH2-ss and
CH3-ss stretching frequency of the hydrocarbon chain of
DPPG lipid increases (Figure 2b,c), whereas the intensity of
CH3-as decreases. It is noticeable that the change in CH3-as
stretching (∼2965 cm−1) of the hydrocarbon of DPPG is
heavily affected by the huge background of the interfacial water
signal.
The VSFG signal of the CH2-ss of the lipid provides
information about the local conformation of the hydrocarbon
chain of the lipid as it appears due to the gauche defects.24,32
The intensity of the terminal CH3-ss signal depends on the
dipole alignment of the hydrocarbon chain of the lipid, which
makes them a sensitive probe of the local orientational order of
the lipid tail.24 Indeed, the intensity ratio of the CH3-ss/CH2-
ss has been utilized to assess the overall order of the lipid
monolayer.32 The addition of dopamine increases the intensity
of CH2-ss of DPPG (Figure 2b), suggesting the increase in the
gauche defects by the local perturbation in the conformation of
the hydrocarbon chain of anionic lipid. The CH3-ss of DPPG
also increases (Figure 2c) on the addition of dopamine,
indicating that the orientational ordering of the hydrocarbon
chain gets enhanced under the effect of dopamine. The
intensity of CH3-ss/CH2-ss decreases (Figure 2d) on the
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addition of dopamine (more specifically at high concen-
tration), which indicates the decrease in the overall ordering of
the hydrocarbon chain of DPPG. The decrease in the overall
ordering of DPPG suggests that increase in the gauche defects
overtakes the effect of the orientational ordering of the
hydrocarbon chain on the addition of high concentration of
dopamine.
The VSFG data of water and C−H stretching regions of
DPPG in the presence of serotonin at the air/DPPG/water
interface show qualitatively the same trend as observed for
dopamine (Figure S2). The intensity of the O−H stretch of
water increases for the concentration ≤50 μM of serotonin,
whereas it decreases significantly with the further addition of
serotonin and finally becomes almost similar to the air/water
interface. The spectral feature of the C−H stretch of DPPG in
the presence of serotonin suggests that the hydrocarbon chain
of DPPG gets disordered at high concentrations of serotonin.
The only difference between the behavior of dopamine and
serotonin at the DPPG interface is that the requirement of
serotonin concentration to affect the water and hydrocarbon
chain of the air/DPPG/water interface is less than dopamine,
which is due to the higher lipophilicity of serotonin than
dopamine.10,11 The VSFG spectra of serotonin at the air/
DPPG/water interface were also measured (Figure S3a) at
physiological pH (7.4, 10 mM Tris-HCl buffer), and the trend
was qualitatively similar to that without buffer conditions (pH
∼5.8). The VSFG study of dopamine at physiological pH
could not be performed because of the oxidation of dopamine
within the measurement time scale.
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Figure 3. (a) SSP polarized VSFG spectra of zwitterionic air/DPPC/water interface on the addition of different concentrations of dopamine. The
changes in the CH3-ss, CH2-ss, and CH3-ss/CH2-ss of the hydrocarbon chain of DPPC for the various concentrations of dopamine are depicted in
panels b, c, and d, respectively. The surface pressure of the DPPC during the measurement was 35 ± 5 mN/m (∼44.5 ± 1.2 Å2/molecule)
depicting the LC phase of lipid. The addition of dopamine does not affect the surface pressure of lipid within the uncertainty of the measurement.

To understand the effect of neurotransmitters on the surface
water near the zwitterionic lipid-containing model membrane,
VSFG studies of the air/DPPC/water interface without and
with dopamine and serotonin have been performed. The VSFG
spectrum of the air/DPPC/water interface is in accordance
with the earlier studies as the intensity of water signal at the
air/DPPC/water interface is higher than that in the air/water
interface (Figure S1c).34,41,46 Irrespective of the zero net
charge on DPPC, it possesses higher dipole moment (∼24 D)
because of the presence of NH3+ and PO4−.47 In earlier studies,
it has been suggested that the high dipole moment of the
DPPC orients the surface water, which results in the higher
VSFG signal of interfacial water at the air/DPPC/water than
the air/water interface.46 In the presence of dopamine, the
intensity of the VSFG signal of water molecules at the air/
DPPC/water interfaces decreases monotonically (Figure 3a),
which indicates that dopamine interacts with the DPPC lipid
irrespective of the fact that it does not possess any net charge.
The decrease in the VSFG signal of interfacial water in the
presence of dopamine suggests that dopamine goes to the
interfacial region near the DPPC and decreases its effective
dipole moment, resulting in the reduction of the oriented water
at the air/DPPC/water interface. The VSFG data indicates
that the NH3+ group of dopamine approaches near the PO4−
group of DPPC lipid at the interface and partially neutralizes
them, which reduces the dipole moment of DPPC lipid.
Indeed, in one of the earlier simulation studies, it has been
suggested that the NH3+ group of neurotransmitters interacts
with the PO4− group of the phosphocholine lipid, which is in
agreement with our VSFG data.16
In agreement with the earlier studies, the SSP polarized
VSFG spectra in the C−H stretching region of DPPC at the
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air/DPPC/water interface show peak features at 2850 and
2883 cm−1 and relatively broad features with a peak at 2950
cm−1. The peak feature at 2850 cm−1 has been attributed to
CH2-ss, whereas the peak features at 2883 and 2950 cm−1 are
assigned to the CH3-ss and CH3-FR with CH3-as of the
hydrocarbon chain of DPPC, respectively.24,40 With the
addition of dopamine, the intensity of CH2-ss and CH3-ss of
the hydrocarbon chain of DPPC increases (Figure 3b,c),
whereas the intensity of the CH3-as decreases, which could be
related with the fluctuation in the background signal of surface
water. The trend of the intensity of CH2-ss and CH3-ss and the
ratio of CH3-ss/CH2-ss of DPPC on the addition of dopamine
is similar to the case of anionic DPPG lipid. The intensity of
CH3-ss/CH2-ss decreases with the addition of dopamine,
especially at the high concentration, suggesting that dopamine
decreases the overall ordering of the hydrocarbon chain of the
zwitterionic DPPC lipid.
Further, the VSFG spectra of the air/DPPC/water interface
were measured without and with serotonin, and the results are
qualitatively similar to those of dopamine (Figure S4). Similar
to dopamine, the NH3+ group of serotonin also reaches near
the phosphate of DPPC and decreases its net dipole moment
which reduces the VSFG signal of water. Serotonin also
decreases the ordering of the hydrocarbon chain of DPPC.
Indeed, earlier thermo-chemical along with MD simulation
studies also concluded that serotonin interacts with zwitter-
ionic lipid vesicles.16 The VSFG spectra of the air/DPPC/
water interface without and with serotonin were also measured
(Figure S3b) at the physiological pH (7.4, Tris-HCl Buffer),
and the trend was qualitatively similar to that without buffer
condition (pH ∼5.8). The VSFG study suggests that the
amine-based chemical neurotransmitters also interact with the
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Figure 4. (a) Probability distribution of different atoms of negatively charged DPPG lipid and dopamine. (b) Snapshot of the intrusion of
dopamine in the DPPG monolayer. The radial distribution functions [g(r)] of phosphate and carbonyl groups of DPPG with different groups of
dopamine are depicted in panels c and d, respectively. g(r) for the interaction of water molecules with different groups of dopamine is depicted in
panel e.

Figure 5. (a) Probability distribution of different atoms of zwitterionic lipid DPPC and dopamine. (b) Snapshot of the intrusion of dopamine in
DPPC monolayer. g(r) of the phosphate and carbonyl groups of DPPC with different groups of dopamine are depicted in panels c and d,
respectively. g(r) for the interaction of water molecules with different groups of dopamine is depicted in panel e.

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Figure 6. Deuterium order parameter (⟨SCD⟩) for DPPG (a) and DPPC (b) monolayer without (black line) and with dopamine (red line).
net neutral zwitterionic lipid-containing model membrane. The
NH3+ of neurotransmitters reaches the interface region of
zwitterionic lipid and interacts specifically with its PO4−
moiety, which reduces the dipole moment of the headgroup
of the zwitterionic lipid, causing decrease in the orientation of
interfacial water along with the decrease in the ordering of the
hydrocarbon chain of the zwitterionic lipid-containing model
membranes. One noticeable observation is that the fraction of
the disordering of the hydrocarbon chain by the neuro-
transmitters is relatively higher for the negatively charged lipid
(∼50%) than the zwitterionic case (∼25%), which probably
suggests that neurotransmitters intrude more into the hydro-
carbon chain of the negatively charged lipid than the
zwitterionic.
In order to get the molecular picture of the interaction of
neurotransmitters with anionic and zwitterionic lipids, MD
simulation studies of DPPG and DPPC lipid monolayer in the
presence of dopamine have been performed for 250 ns using
the GROMACS 2016.348 package (the detailed simulation
protocol is described in the Supporting Information). The
similarity of the experimental data of dopamine and serotonin
with lipids suggests that the mode of interaction of serotonin
with lipids should be similar to that of dopamine; hence, only
dopamine was chosen for the simulation. The simulated
average lateral area per lipid for DPPG and DPPC was found
to be ∼55 ± 2 and ∼48 ± 2 Å2, respectively (Figure S5a,b),
which is in line with the experimental condition as mentioned
in the captions of Figures 2 and 3. The simulated data also
show that the addition of dopamine does not affect the average
lateral area per lipid, which is in line with experimental
observation. The structural properties of anionic and
zwitterionic phospholipid monolayer and the adsorption
behavior of the neurotransmitter were elicited using the
positional probability distributions of the selected atoms of
lipids and dopamine [DPPG and DPPC lipids: phosphorus
atom of the phosphate group (P), the carbonyl group (C1,
glycerol backbone), terminal methyl group (C16), the nitrogen
of the choline of DPPC(N), the oxygen atom of water (Ow);
dopamine: OH and OH1, NH3+ groups] as shown in panels a
and b of Figure 4, respectively. The positional probability
distribution data of the lipid monolayer system suggest the
broad distribution of the polar headgroup (P for DPPG, P and
N for DPPC), a flexible hydrocarbon tail, as well as the
incursion of water into the lipid headgroup region consisting of
polar groups in addition to glycerol regions, which resemble
the results of earlier reports of the bilayer of phospholipids.12,16
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The simulation data depict that dopamine intrudes into both
lipid monolayers as the probability of dopamine is high in the
headgroup region of DPPG as well as DPPC, although the
intrusion of dopamine is greater in DPPG than DDPC. It can
be seen that the probability of the existence of the OH group
of dopamine is higher near the carbonyl group, whereas the
NH3+ group of dopamine locates between the phosphate and
carbonyl group of DPPG and DPPC lipids, which is also
supported by the snapshots (Figures 4b and 5b).
To further quantify the interaction between DPPG lipid and
dopamine, the radial distribution function [g(r)] between the
different groups of the headgroup region of phospholipid and
dopamine has been calculated, as shown in Figure 4c−e. The
g(r) data suggest that both NH3+ and OH groups of dopamine
may interact with the PO4− of DPPG; however, the interaction
of the NH3+ with the PO4− headgroup is significantly higher
than the OH group (Figure 4c). Conversely, the OH group of
dopamine interacts significantly with the carbonyl group of the
lipid (Figure 4d). The prominent interaction of NH3+ and OH
groups of dopamine with PO4− and carbonyl headgroup of
lipid is also supported by the probability distribution as well as
the snapshot feature, which depicts that the NH3+ group of
dopamine is mainly pointed down toward the phosphate of
DPPG, whereas OH is pointed up toward the carbonyl group.
The g(r) of the interaction of dopamine with water suggests
that both NH3+ and OH groups of dopamine interact with
water molecules present in the headgroup of the DPPG lipid
(Figure 4e). However, the interaction of dopamine with
interfacial lipid water is significantly less than its interaction
with the DPPG lipid. Further, the probability distribution of
the oriented water near the phosphate group of DPPG without
and with dopamine has been calculated (Figure S6b). The data
suggest that the hydrogen of water is up-oriented (∼130°)
near the phosphate group of DPPG, and the probability of up-
oriented water decreases away from the interface as the
randomization of water takes place in the bulk medium. Upon
the addition of dopamine, the orientation of water near the
phosphate group is similar to the pure DPPG case (Figure
S6b). However, the probability of up-oriented water decreases
slightly in the vicinity of the PO4− group of DPPG in the
presence of dopamine, suggesting the decrease of the probe
length of interfacial water in line with the experimental finding.
The simulation data of DPPG along with the experimental data
indicate that the NH3+ group of dopamine intrudes into the
lipid by removing the surface lipid water and interacts with the
phosphate group of DPPG.
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The snapshots (Figure 5b), as well as g(r) data of DPPC and
dopamine (Figure 5c,d), suggest that the NH3+ of dopamine
prefers to predominantly interact with PO4− of DPPC, whereas
OH of dopamine is accommodated preferably near the
carbonyl group of the zwitterionic lipid. The interaction of
the OH of dopamine with the choline group of DPPC is very
weak as the g(r) for this interaction is broad and appears at
greater distance (Figure 5c). The g(r) data of water indicates
that both NH3+ and OH groups of dopamine interact with
water. However, dopamine prefers to interact mainly with the
lipid headgroup as compared to interfacial lipid water
molecules (Figure 5e). In comparison to the DPPG case, the
decrease in the orientation distribution of water in the vicinity
of the phosphate of DPPC is significantly less (Figure S6c),
which could be due to the different nature of the headgroup of
DPPG and DPPC. Indeed, the decrease in the VSFG signal of
water is higher in DPPG than DPPC upon the addition of
dopamine in the experimental data also. The experimental and
simulation data both suggest that the NH3+ group of dopamine
gets closer to the phosphate of the DPPC. The possible
interaction of the NH3+ group of dopamine with the phosphate
of DPPC decreases the dipole moment of the headgroup of
DPPC, resulting in less oriented interfacial lipid water.
Further, the deuterium order parameter (⟨SCD⟩) of the
hydrocarbon chains DPPG and DPPC without and with
dopamine have been calculated to get insight into the effect of
a neurotransmitter on the ordering of the hydrocarbon chains
of the lipid monolayer (Figure 6).49,50 It is apparent that the
order parameter of the hydrocarbon chain of both lipids
decreases in the presence of dopamine, indicating that the
hydrocarbon chain of both lipids gets disordered in the
presence of dopamine, which is in line with the experimental
observations discussed above. Indeed, the tilt angle (the angle
between the monolayer normal and the directive vector of the
alkyl chains) distribution of the hydrocarbon chain of both
lipids (Figure S7a−c) increases with the addition of dopamine,
which also suggests the disordering of lipids induced by
dopamine. It is well-known that the hydrophobic interaction
between the hydrocarbon chains plays a key role in its
ordering.50 The g(r) data for the interaction of hydrocarbon
chains of both lipids decrease in the presence of dopamine in
comparison to that of pure cases (Figure S8). The data also
suggest that the interaction of dopamine with hydrocarbon
chains is not significant, suggesting that the direct interaction
of dopamine with the hydrocarbon chains of both lipids is not
significant as compared to the interaction between the
hydrocarbon chains (Figure S8). Hence, the decrease of the
ordering of chains of lipids upon the addition of dopamine is
mainly due to the decrease in the interaction between their
hydrocarbon chains.
The direct interaction of chemical neurotransmitters,
namely, dopamine and serotonin, with anionic and zwitterionic
phospholipid monolayers was explored utilizing the surface-
specific vibrational spectroscopy in the C−H and O−H
stretching regions. The intensity of the water signal decreases
in the presence of higher concentration of dopamine,
suggesting the intrusion of neurotransmitters into the head-
group of the anionic lipid monolayer where the NH3+ group of
neurotransmitters interacts with the phosphate moiety of the
DPPG lipid. Neurotransmitters also intrude into the net
neutral zwitterionic DPPC lipid. The NH3+ groups of
neurotransmitters specifically interact with the phosphate
moiety of DPPC, which decreases the orientation of interfacial
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water created by the dipole of the headgroup of DPPC. The
intrusion of neurotransmitters into the headgroup regions of
both anionic and zwitterionic lipids decreases the interaction
between the hydrocarbon chains of lipids and changes its local
conformation, which leads to the disordering of the hydro-
carbon chain of lipids. The VSFG and MD simulation studies
indicate that the direct interaction of the neurotransmitters
significantly restructures the membrane water as well as the
ordering of phospholipids, which can be utilized to understand
the complex synaptic signaling processes.
■
*
ASSOCIATED CONTENT
sı Supporting Information
The Supporting Information is available free of charge at
https://pubs.acs.org/doi/10.1021/acs.jpclett.1c00173.
Experimental methods, molecular dynamics simulation
protocol, and supporting figures (PDF)
■ AUTHOR INFORMATION
Corresponding Author
Prashant Chandra Singh − School of Chemical Sciences,
Indian Association for the Cultivation of Sciences, Jadavpur,
Kolkata 700032, India; orcid.org/0000-0001-9357-
9039; Email: sppcs@iacs.res.in
Author
Biswajit Biswas − School of Chemical Sciences, Indian
Association for the Cultivation of Sciences, Jadavpur, Kolkata
700032, India; orcid.org/0000-0002-9075-1689
Complete contact information is available at:
https://pubs.acs.org/10.1021/acs.jpclett.1c00173
Notes
The authors declare no competing financial interest.
■
ACKNOWLEDGMENTS
The financial support from DST-SERB India (Grant Number:
CRG/2019/001389) and CRAY for computation of IACS are
gratefully acknowledged. B.B. thanks CSIR and IACS for
fellowship.
■
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