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How to cite: Angew. Chem. Int. Ed. 2020, 59, 22731 – 22737
Interfaces International Edition: doi.org/10.1002/anie.202011179
German Edition: doi.org/10.1002/ange.202011179

Interaction of a-Synuclein with Phospholipids and the Associated

Restructuring of Interfacial Lipid Water: An Interface-Selective
Vibrational Spectroscopic Study
Biswajit Biswas, Subhadip Roy, Jahur Alam Mondal,* and Prashant Chandra Singh*

Abstract: Interaction of a-Synuclein (aS) with biological
lipids is crucial for the onset of its fibrillation at the cell
membrane/water interface. Probed herein is the interaction of
aS with membrane-mimicking lipid monolayer/water interfa-
ces. The results depict that aS interacts negligibly with
zwitterionic lipids, but strongly affects the pristine air/water
and charged lipid/water interfaces by perturbing the structure
and orientation of the interfacial water. The net negative aS
(@9 in bulk water; pH 7.4) reorients the water as hydrogen-up
(H-up) at the air/water interface, and electrostatically interacts
with positively charged lipids, making the interface nearly net
neutral. aS also interacts with negatively charged lipids: the net
H-up orientation of the interfacial water decreases at the
anionic lipid/water interface, revealing a domain-specific
interaction of net negative aS with the negatively charged
lipids at the membrane surface.
Introduction
(@12) and hydrophilic; whereas the NAC region is highly
hydrophobic with only one unit negative charge (@1). At the
physiological concentration ( & 50 mM), aS does not aggre-
gate in bulk aqueous phase (pH & 7.4), even on keeping the
solution for several days.[7] In contrast, aS exhibits accelerated
aggregation at membrane-water interfaces (bulk pH
& 7.4).[8–10] Aggregation proceeds via pre-nucleation which is
essentially the anchoring of soluble monomeric aS with
membrane lipid head groups at the interface. Pre-nucleation
controls the subsequent primary and secondary nucleation
processes (conformation change) where different sized nuclei
of aS are formed, leading to the formation of fibrils.[9, 11] The
conformational change of disordered protein at membrane-
water interface has been demonstrated using the chiral-VSFG
technique.[12, 13] Several studies have suggested that the nature
of lipid as well as the lipid:aS ratio controls the rate of
aggregation.[7–9, 11] For example, aggregation is more pro-
nounced at the surface of negatively charged lipids than that

ParkinsonQs and AlzheimerQs diseases are the two impor-

tant neurodegenerative disorders associated with progressive
dementia. The appearance of amyloid plaques in the brain is
the hallmark of such disorders, which in turn linked with the
aggregation of misfolded cellular proteins followed by
fibrillization.[1–3] Hence, understanding the aggregation of
misfolded protein is indispensable for early intervention to
amyloid plaque formation.
a-Synuclein (aS; 140 amino acid residues; Figure 1 a is
one of the disordered proteins whose aggregation is corre-
lated with ParkinsonQs and Lewy body diseases.[4, 5] As shown
in Figure 1 a, the primary structure of aS contains three main
domains, namely N-terminal (blue color), non-amyloid b
component (NAC, ash color), and C-terminal (red color).[6]
The N-terminal contains four units positive charge (+ 4) and
amphipathic in nature; the C-terminal is negatively charged

[*] B. Biswas, Dr. P. C. Singh

School of Chemical Sciences
Indian Association for the Cultivation of Sciences
2A &2B Raja S. C. Mullick Road, Jadavpur, Kolkata 700032 (India)
E-mail: sppcs@iacs.res.in
S. Roy, Dr. J. A. Mondal
Radiation & Photochemistry Division, Bhabha Atomic Research
Centre, Homi Bhabha National Institute Figure 1. a) Amino-acid sequence of aS and the regions of N-terminal,
Trombay, Mumbai 400085 (India) NAC, and C-terminal. Negatively charged residues are shown in red,
E-mail: mondal@barc.gov.in whereas positively charged residues are in blue. b) Chemical structure
Supporting information and the ORCID identification number(s) for of the lipids used: positively charged DPTAP, negatively charged DPPG
the author(s) of this article can be found under: and DPPS, and zwitterionic DPPC. The counter ions with the charged
https://doi.org/10.1002/anie.202011179. lipids are omitted for simplicity.

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Research Articles
of the positively charged and zwitterionic lipids. For a fixed
concentration of monomeric aS, the aggregation increases
linearly with the concentration of negatively charged lip-
id.[9, 14] However, it is intriguing to observe that a negatively
charged lipid surface promotes the aggregation of a net
negatively charged (@9) protein which is supposed to be
repelled from the membrane surface.
Apart from lipid head groups, interfacial water is believed
to be crucial for the aggregation process. Time-resolved
fluorescence measurement showed slower solvation of water
near negatively charged surfactant head group which pro-
motes fibrillization of disordered protein.[15, 16] MD simulation
combined with time-resolved fluorescence suggests longer
trapping of interfacial water (slower solvation) near the
amyloidogenic region that triggers pathological conversion of
aS into amyloid form.[17] “Electron” and “atomic force”
microscopy measurements also reveal that the air–water
interface affects the pre-nucleation and aggregation of aS.[18]
Undoubtedly, the aggregation of aS at membrane-water
interface is a complex interplay of lipid-protein, lipid-water,
protein-water, and water-water interactions. Conventional
spectroscopic studies, such as circular dichroism (CD),[9]
molecular probe-based fluorescence lifetime and microsco-
py,[18, 19] provide information about the changing rate of aS
aggregation at membrane surface, but the role of interfacial
water and the molecular origin of lipid head group-specificity
towards the accelerated aggregation are not well understood.
Interface-selective spectroscopy such as vibrational sum
frequency generation (VSFG) can provide molecular level
insight into the restructuring of interfacial water and the
associated lipid-protein interaction during pre-nucleation of
aS at model membrane-water interfaces. Understanding the
pre-nucleation at membrane-interface is important as the
monomeric aS interacts with the lipid head group during this
stage which leads to the primary and secondary nucleation for
the fibrillation process.[9, 11] VSFG spectroscopy has been
extensively used to unravel water structure and protein
conformational changes at aqueous interfaces.[20–23] Further-
more, heterodyne detection of VSFG signal (heterodyne-
detected VSFG), enabled us to directly measure the preferred
orientation of interfacial molecules through the sign of Imc(2),
which is particularly advantageous in pinpointing the specific
interaction between charged groups at the interface.[24–27]
Figure 2. a) Imc(2) spectra of the neat air–water and air–water-aS interfaces (surface pressure, p = 20 : 3 mN m@1); Inset: expanded view of the
OH stretch region. b) Imc(2) spectra of cationic lipid (DPTAP)-water and DPTAP-water-aS interfaces (p = 35 : 3 mN m@1).
22732 www.angewandte.org T 2020 Wiley-VCH GmbH
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Previous HD-VSFG studies by us and others have shown
unique orientation of water at different lipid-water interfaces
according to the charge of the lipid head groups.[28–33]
In this article, we clarified the lipid head group specificity
towards the interaction with aS and the associated reorgan-
ization of interfacial water using surface tensiometry and
broadband HD-VSFG measurements. The air–water and
various lipid monolayer-water interfaces (e.g., cationic 1,2-
dipalmitoyl-3-trimethylammonium propane chloride salt
(DPTAP), anionic 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-
1-glycerol sodium salt (DPPG), 1, 2-dipalmitoyl-sn-glycero-3-
phosphoserine sodium salt (DPPS) as well as zwitterionic 1,2-
dipalmitoyl-sn-glycero-3-phosphocholine (DPPC); (Fig-
ure 1 b) were measured in the presence and absence of aS
in the respective aqueous phases. The Imc(2) spectra (OH and
CH stretch regions) display significant effect of aS at the air–
water and charged lipid-water interfaces: the “H-down”
oriented water becomes “H-up” in presence of aS (20 mM) at
air/water interface, signifying the accumulation of aS at the
air–water interface that makes the interface negatively
charged. At the lipid monolayer-water interface, aS is
electrostatically attracted to the charged lipid head groups;
however, it hardly interacts with the zwitterionic lipid (net
neutral) head group. Specifically, at the cationic DPTAP-
water interface, the H-down orientation of interfacial water
decreases in the presence of aS (20–50 mM) due to screening
of the lipid positive charge. In the case of anionic DPPG- and
DPPS-water interfaces, aS (20–50 mM) moderately decreases
the net H-up orientation of interfacial water, suggesting the
interaction of positively charged N-terminal of aS with
negatively charged lipid head groups. Thus, despite being
net negative (@9), aS attractively interacts with anionic lipids
via its N-terminal at the membrane-water interface.
Results and Discussion
Figure 2 a shows the Imc(2) spectra (2660–3660 cm@1) of
the neat air–water and air–water-aS (20 and 50 mM) inter-
faces. The Imc(2) spectrum of the air–water interface (black
dashed curve) shows a broad negative band, centered around
3450 cm@1, and a positive band above & 3580 cm@1. In agree-
ment with previous reports,[34–36] the negative Imc(2) band is
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due to “H-down” orientated interfacial water; whereas the
positive band above 3580 cm@1 whose maximum is
& 3700 cm@1 (not clearly visible in the present measurement
due to insufficient IR power in that region) is due to “H-up”
orientated dangling OH of the topmost water. It is important
to note that previous phase-sensitive/heterodyne-detected
VSFG measurements[20, 27, 37, 38] showed a positive Imc(2) band
around 3200 cm@1 and was assigned to the “ice-like” or
“strongly H-bonded water pair”. The positive band however
is not so prominent in the carefully measured recent Imc(2)
spectra,[35, 36, 39–41] which is in conformity with the present data
(black-dashed curve in Figure 2 a ) measured using high
resolution displacement sensor (see the Supporting Informa-
tion) as well as our previously reported spectra that were
phase-corrected by internal referencing.[42–44] With the addi-
tion of 20 mM aS in water, the surface pressure (p = surface
tension of neat water (72.8 mN m@1)—surface tension of aS
solution) becomes & 20 mN m@1, indicating adsorption of aS
on water surface. In the HD-VSFG measurement too, the CH
stretch region (< 3000 cm@1) which was featureless for the
neat air–water interface, shows sharp negative bands around
2880 and 2940 cm@1 (yellow curve) corresponding to the C@H
stretch of aS, confirming the presence of aS at the air–water
interface. The OH stretch band ( & 3000–3580 cm@1) becomes
positive in sign due to interfacial aS (yellow curve, expanded
view of the OH stretch is shown in the inset, Figure 2 a). The
Positive sign implies that the interfacial water which was H-
down oriented (negative Imc(2)) at the neat air–water inter-
face, adopts H-up orientation in presence of aS. At pH 7.4,
monomeric aS contains both positively and negatively
charged domains (Figure 1 a) however, the net charge is
negative (@9). Thus, it is the net charge of aS that makes the
interface negatively charged and preferentially orients the
interfacial water as H-up (positive Imc(2)), though it is not
obvious whether the net charge on the interfacial aS is @9 or
less presumably due to the modified protonation Ð depro-
tonation equilibrium at the interface. A cursory glance to the
OH stretch band (at air–water-aS) reveals that its amplitude
is significantly lower than that of charged lipid-water inter-
faces (compare with the solid curves in Figure 2 a and 3 a). It
can be concluded that the adsorption of aS makes the
interface only weakly negatively charged, which is either due
to partial protonation of aS at the interface or due to the
immersion of the negatively charged strongly hydrated
carboxyl groups deep into the aqueous phase such that they
contribute negligibly to the net orientation of water at the
interface. The dip feature observed around 3280 cm@1 (yellow
curve, Figure 2 a) is presumably due to the N-H stretch of
protein as observed in the chiral-VSFG study.[13] In the region
above 3580 cm@1, the positive signal though noisy is distinctly
observable in the presence of aS (20 mM).
With the increasing bulk concentration of aS (50 mM; blue
curve, Figure 2 a), the Imc(2) signal increases in the CH stretch
region, suggesting a higher number density of aS at the
interface. To avoid aggregation in bulk water, we did not
increase aS concentration beyond 50 mM.[9, 45] Since aS is net
negative (@9; pH 7.4), its increased number density is
expected to enhance negative charge density at the interface,
augmenting the H-up orientation of interfacial water already
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present at the 20 mM aS solution. However, the positive Imc(2)
signal in the OH stretch region (3000–3500 cm@1) decreases,
though marginally. It indicates reduced orientational prefer-
ence of interfacial water at higher concentration of aS
(50 mM). The decreased water signal suggests a possible
conformational change of aS at the air–water interface such
that the relative influence of the positively charged hydro-
phobic N-terminal of aS increases on interfacial water. This
proposition agrees with recent electron- and atomic force
microscopic measurements that suggested a conducive role of
the aqueous interface towards the primary and secondary
nucleation and aggregation of aS.[18] The positive band around
3600 cm@1 is clearly visible even for 50 mM aS, pointing to the
existence of H-up oriented weakly interacting OH at the air–
water-aS interface. Given the hydrophobic nature of the N-
terminal, the existence of such weakly interacting water is
quite likely at the air–water–aS interface.[28, 46] We note that
the peak position of the OH stretch band appears around
3200 cm@1 in the presence of aS, which is red-shifted from that
of the neat air–water interface ( & 3450 cm@1). The apparent
red-shift may not be solely due to stronger H-bonding of
interfacial water; the spectral deformation due to vibrational
coupling of interfacial water[47] and the reversal of the sign of
Imc(2) in presence of aS may also contribute to the apparent
maximum around 3200 cm@1.
Next, we measure the Imc(2) spectra (Figure 2 b) of the
cationic DPTAP-water interface in the presence and absence
of aS (20 and 50 mM). The Imc(2) spectrum of the DPTAP-
water interface (solid black curve) shows a negative double-
peak feature in the CH stretch region corresponding to the
terminal methyl symmetric stretch (CH3-SS, 2880 cm@1) and
Fermi resonance with its bend overtone (CH3-FR,
2940 cm@1).[48] The CH3-SS band appears with a negative sign
due to “CH3-up” orientation (i.e., methyl hydrogens are
pointed towards the air or away from the aqueous phase) of
DPTAP alkyl chains on the water surface.[27] A weak shoulder
band around 2850 cm@1 corresponds to the methylene sym-
metric stretch (CH2-SS) of the alkyl chains. Although, the
number density of -CH2- is higher than that of -CH3, the CH2-
SS signal is much weaker than CH3-SS due to local inversion
symmetry in the well-packed alkyl chains of DPTAP mono-
layer.[49] In the high frequency region of the CH stretch,
a positive band is observed around 2965 cm@1, corresponding
to the CH3-AS of the terminal methyl of the lipid. The C@H
stretch signal of DPTAP remains almost invariant with 20 mM
aS, but decreases slightly on increasing the concentration of
aS to 50 mM.
The OH stretch signal at the DPTAP-water interface is
significantly higher than the neat air–water interface (com-
pare solid- and dashed-black curves in Figure 2 b) due to
increased preferential H-down orientation of water at the
cationic lipid interface following the principle of electro-
statics. The Imc(2) spectrum shows a strong negative band in
3000–3570 cm@1 regions, followed by a weak positive band
above 3570 cm@1. The intense negative band is due to the
large population of H-down oriented water in the electric
double layer (EDL) below the positively charged lipid
monolayer; while the weak positive band is due to weakly
interacting H-up oriented water in the glycerol region of the
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lipid.[28, 32] It is worth noting that the opposite orientations of
CH3-SS and OH-SS (methyl-up vs. H-down) corresponding to
the same negative sign of Imc(2) is due to the opposite signs of
the hyperpolarizability (b) of CH3-SS and OH-SS vibrational
modes.[27]
With the addition of 20 mM aS, the negative Imc(2) signal
(OH stretch; red curve) decreases and the central frequency
of the band shifts towards higher frequency. This result clearly
suggests that the net negative aS screens the positive electric
field of the DPTAP monolayer in the interfacial region such
that thickness of the EDL containing the preferentially H-
down oriented water decreases. On increasing the concen-
tration of aS to 50 mM, the negative OH stretch signal
decreases to such an extent that it becomes comparable to
that of the pristine air–water interface (compare solid-green
and dashed-black curves, Figure 2 b). With the higher con-
centration of aS (50 mM) the observed spectral changes at the
DPTAP-water interface are instantaneous, while those at the
lower concentration of aS (20 mM) show modest time-
dependence (see Figure S4 in the Supporting Information).
The Imc(2) spectra shown in Figure 2 and subsequently are
equilibrated spectra recorded after a waiting time of several
hours. The spectral changes in Figure 2 b clearly show that the
negatively charged aS is electrostatically attracted towards
the positively charged head group of DPTAP such that the
water in the interfacial region does not experience any net
electric field (i.e., similar to the uncharged air–water inter-
face) at 50 mM bulk concentration of aS. In other words, the
aS completely screens the positive charge of the lipid head
group and the corresponding Imc(2) signal in 3100–3650 cm@1
region selectively represents the response of the interfacial
water free from the bulk-like oriented water in the EDL[50, 51]
at the DPTAP-water-aS interface. The blue-shift of the
central frequency of the Imc(2) spectra (OH stretch) suggests
that the vibrational coupling and/or H-bonding of interfacial
water decrease in presence of aS at the cationic DPTAP-
water interface.
The Imc(2) spectrum of the negatively charged DPPG-
water interface (black curve Figure 3 a) shows two negative
and one positive band in the CH stretch region, corresponding
to the CH3-SS, CH3-FR, and CH3-AS bands, similar to that of
the DPTAP-water interface (Figure 2 b). Presence of aS
Figure 3. a) Imc(2) spectra of anionic DPPG-water and DPPG-water-aS interfaces (p = 35 : 3 mN m@1). b) Imc(2) spectra of net negative DPPS-
water and DPPS-water-aS interfaces (p = 35 : 3 mN m@1). The spectrum of the neat air–water interface (dashed curve) is shown in each panel for
reference.
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marginally changes the amplitudes of the CH stretch bands.
In the OH stretch region ( & 3000–3650 cm@1), the Imc(2)
spectrum is positive in sign and high in amplitude due to
preferred H-up orientation of water at the negatively charged
DPPG interface and the associated EDL region.[29, 32, 33]
Presence of aS (20 mM) slightly decreases the positive Imc(2)
signal (compare red and black curves, Figure 3 a). On
increasing the concentration of aS (50 mM), the positive OH
stretch signal decreases further, but not as significant as that
of the positively charged DPTAP interface. The decreased
positive Imc(2) signal is a manifestation of the reduced
negative surface electric field at the DPPG-monolayer in
the presence of aS, presumably via the attractive interaction
between the anionic lipid headgroup and the positively
charged N-terminal of aS. In the case of another negatively
charged lipid, DPPS, similar spectral changes are observed in
the presence of 20 and 50 mM aS (Figure 3 b). DPPS has the
same net charge (@1) as that on DPPG, but the total number
of charged moieties in its head group are different (PO4@ ,
COO@ and NH3+; Figure 1 b). From the perspective of
interfacial water, similar spectral changes at the DPPG and
DPPS interfaces confirm that aS interacts with net negative
lipids with equal efficiency even when the lipids have
a different distribution of charge moieties in their head
groups. This observation confirms that the lipid-aS interac-
tion is electrostatic in nature.
For further insight into the interaction, we performed MD
simulation of DPPG-water and DPPG-water-aS systems (see
the Supporting Information). The positional probability
calculation (Figure S7b) suggests an overlapping region
between the phosphate (PO4@) group of DPPG and aS, in
particular, with the positively charged lysine (LYS) at the N-
terminal of aS. The radial distribution functions (RDFs)
between PO4@ of DPPG and LYS (or aS) shows a strong
interaction between LYS and PO4@ (Figure S7c), reinforcing
the notion that the positively charged LYS at the N-terminal
of aS is electrostatically attracted towards the DPPG head-
group. It is important to mention that we did not find any
significant overlap between the hydrocarbon chain of DPPG
and aS, suggesting that the interaction is localized within the
aqueous interfacial region without a noticeable effect on the
lipid hydrophobic region. Orientational distributions of water
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depict H-up orientation of water at the DPPG-water at
DPPG-water-aS systems (Figure S8), which is in line with the
experimental results. However, unlike DPPG, water near to
the aS have both the “H-up” and “H-down” orientations,
which could be due to the contribution of different local
amino acids of aS.
In addition to the charged lipids, we measured the
interaction of aS with zwitterionic DPPC lipid (Figure 4).
Imc(2) spectrum of the DPPC-water interface (in absence of
aS) shows typical CH stretch bands below 3000 cm@1,
followed by a positive band around 3200 cm@1 and a dip-
feature around 3450 cm@1. The 3200 cm@1 band is mainly due
to anionic phosphate associated water while the dip-feature
around 3450 cm@1 is predominantly due to the cationic
choline associated water.[30] Presence of aS (up to 50 mM)
hardly affects the Imc(2) spectrum at the DPPC-water inter-
face, which is a manifestation of negligible interaction of aS
with the zwitterionic DPPC. This observation in combination
with the results at the charged lipid interfaces further suggests
that the hydrophobic alkyl chains of the lipids do not directly
interact with aS; it is mainly the lipid headgroups that interact
electrostatically with the charged domains of aS. This
observation qualitatively agrees with the less pronounced
aggregation of aS at zwitterionic lipid vesicles than that of the
negatively charged lipids.[8, 9] Since, many laboratories, instead
of Imc(2), report the SFG-intensity i.e., j c(2) j 2-spectra, we
showed the deduced j c(2) j 2-spectra corresponding to Imc(2)
spectra of Figure 2 b, 3a and 4, in the Supporting Information
(Figure S5).
In summary, our results reveal that aS effectively interacts
with those lipids that have charged head groups, and the
interaction is primarily electrostatic in nature. The net
negative aS (@9 in bulk water, pH 7.4) is attracted to the
cationic lipid headgroups, which drastically screens the
positive charge and reduces the thickness of the EDL in the
vicinity of the corresponding membrane surface. However,
the most intriguing observation is the screening of the lipid
negative charge (DPPG and DPPS) by the net negative aS.
Anionic lipids are expected to repel the negatively charged aS
(Figure 1 a) such that the interface will be depleted of aS.
Figure 4. Imc(2) spectra of the zwitterionic DPPC-water (black) and
DPPC-water-aS interfaces (red and green; p = 35 : 3 mN m@1). The
spectrum of the neat air–water interface (dashed curve) is shown as
reference.
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Accordingly, the H-up oriented interfacial water that is, the
positive Imc(2) signal (OH stretch) is expected to remain
unaffected (if not increased) in the presence of aS at the
DPPG- and DPPS-water interfaces. However, the experi-
mental results (Figure 3 a and b) show that the positive Imc(2)
signal (OH stretch) decrease in presence of aS at these
interfaces, revealing the screening of the lipid head group
negative charge. We propose that the positively charged N-
terminal of aS preferentially interacts with the negatively
charged head group of DPPG (or DPPS) such that the net
negative aS exhibits an attractive interaction with the
negatively charged membrane headgroups and screens the
negative electric field to some extent in the interfacial region.
Nevertheless, the significant positive OH stretch signal at the
DPPG and DPPS interfaces, even in presence of 50 mM aS,
indicates that the surface field is still strong enough to
preferentially orient the interfacial water as H-up. Currently,
it is not clear whether the negative electric field originates
from the inherent negative charge of the lipid headgroup or
the interfacial aS (net negative) interacting with the lipid
headgroup via its positive N-terminal. Irrespective of the
origin of the net negative charge, it is confirmed that aS
interacts with anionic lipid monolayer via electrostatic
interaction, though from the perspective of the interfacial
water the screening effect of aS on the negative surface field
(DPPG and DPPS interface) is much weaker than that on the
positive surface field (DPTAP interface). This observation is
in conformity with the findings of several non-interface-
selective spectroscopic investigations. For instance, the fluo-
rescence energy transfer study of aS in negatively charged
SDS micelles suggests compaction of N-terminal and expan-
sion of C-terminal regions, leading to the conformational
change of aS.[52] NMR[53] and EPR[54, 55] studies of aS with
synthetic lipid vesicles and detergent micelles also pointed out
that the C-terminal of aS remains free whereas the N-
terminal binds to the vesicle and micelle surfaces, which
triggers fibrillation. Fibril monomer equilibration study of aS
with the varying charge on the C-terminal suggests that
fibrillation is favored for the C-terminal of lower negative
charge.[56, 57] Similar observation of decrease in the aggrega-
tion of aS by its C-terminus is also reported in microscopy,
circular dichroism and FT-IR studies.[58, 59]
Conclusion
We probed the interaction of aS with cationic, anionic,
and zwitterionic lipid monolayers on the water surface by
using interface-selective heterodyne-detected vibrational
sum-frequency generation (HD-VSFG) spectroscopy in the
OH and CH stretch regions. The Imc(2) spectra of the air–
water-aS interface show that the aS adsorbs at the air–water
interface and invert the net orientation of interfacial water to
H-up (i.e., water Hs are pointed towards the air and away
from the aqueous phase), corresponding to a weakly neg-
atively charged aqueous interface. The changes in the Imc(2)
spectra exposes the strong interaction of aS with the
positively charged lipid (DPTAP)-water interface, making
the interface net neutral from the perspective of the
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interfacial water. The net negative aS also interacts with the
anionic lipid (DPPG and DPPS) headgroups presumably via
the positively charged N-terminal of aS; though the effective
screening of the interfacial electric field is much weaker than
that at the cationic lipid-water interface. In the case of
zwitterionic lipid (DPPC), aS does not produce noticeable
changes in the CH and OH stretch regions, suggesting no
preference of aS for the zwitterionic lipid-water interface.
Altogether, the Imc(2) spectral changes suggest that aS
interacts with the charged lipid head groups at the aqueous
interface and the hydrophobic alkyl chains of the lipids
remain largely unperturbed. The aS-lipid interaction and the
associated change in water structure and orientation may play
a crucial role for the fibrillation of aS at the membrane
surface.
Acknowledgements
PCS acknowledges DST-SERB-India [Grant Number: CRG/
2019/001389] for financial support. JAM gratefully acknowl-
edges the support and encouragement from Dr. Awadhesh
Kumar, Head, RPCD and Dr. A. K. Tyagi, Associate
Director, Chemistry Group, BARC. BB thanks Dr. Sampali
Banerjee and Dr. Prosenjit SenQs lab of IACS for help in
protein purification. BB and SR thank CSIR and HBNI,
BARC for their respective fellowship.
Conflict of interest
The authors declare no conflict of interest.
Keywords: electrostatic interactions · lipids · interfaces ·
membranes · Parkinson’s disease
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Manuscript received: August 15, 2020
Revised manuscript received: August 27, 2020
Accepted manuscript online: August 31, 2020
Version of record online: October 7, 2020
www.angewandte.org 22737