T.

Paldi: Salt-Bridge Switching

Salt-Bridge Switching Underlies Membrane

Voltage Sensing

Tzur Paldi

© 2012 Tzur Paldi, All Rights Reserved

1
 T. Paldi: Salt-Bridge Switching
Salt-Bridge Switching Underlies Membrane Voltage Sensing
Tzur Paldi
Abstract Voltage sensing modules (VSMs) are specialized
membrane proteins that change their conformation in
response to changes in the electric potential across the cell
membrane. The VSM is composed of four transmembrane
helix segments (S1-S4), and its most striking feature is a
conserved repetitive motif of a positively charged amino acid
residue followed by two hydrophobic residues, within the
fourth transmembrane segment (S4). It is generally accepted
that the movement of the positively charged residues in the
membrane electric field drives S4, relative to S1-S3.
However, recent study showing the titratable nature of S4
arginines suggests that these arginines are not permanently
charged, and changes in their ionization state may participate
in the VSM voltage-sensing mechanism (Paldi, 2012
http://sdrv.ms/13hN4Vm). Here it is shown that the gating
properties of the bacterial Na+ channel fit a model in which
S4 arginines, salt bridged to acidic residues, are neutralized
via voltage-dependent pKa shifts. A compatible mechanistic
model that simulates the electrical properties of the VSM as
those of a two-plate capacitor, shows that reorientation of a
dipole moment, probably of water molecules, in the low
dielectric septum of the VSM mediates these pKa shifts. The
proposed model demonstrates the role of salt bridge
formation in driving S4 and in preventing ion leaks (“omega”
currents) through the VSM vestibule during S4 movement,
while generating a capacitive gating current by means of
proton relocation. This model agrees with major experimental
aspects, and further reconciles them with the helical screw
mechanism for S4 movement.
Keywords Electric field ∙ Electrostatic interactions ∙
NaChBac ∙ pKa shift ∙ Voltage-dependent ion channels ∙
Voltage sensor domain
On the cover: The evolvement of the voltage sensor module
‘ratcheted capacitor’ theory
__________________________________________________
Author: Tzur Paldi
Email: tzur.paldi@outlook.com
© 2012 Tzur Paldi, All Rights Reserved
__________________________________________________
2
Introduction
Voltage sensing across membranes is fundamental to many
cell functions. One of the most well-known of the membrane
proteins involved in such sensing is the S4-based voltage
sensing module (VSM), which changes its conformation in
response to changes in the electrical potential across the
membrane [1]. VSMs are mostly known for their role in
voltage-gated ion channels in excitable cells and tissues [2].
However, non pore-attached VSMs that are either coupled to
phosphatase [3], or that constitute a proton channel as an
independent module [4, 5], have recently been described.
VSMs are composed of four membrane-spanning helices
(S1-S4). The forth transmembrane segment, S4, is typically
embedded with up to eight positively charged residues at
every third position, while the four outermost arginines (R1-
R4) contribute the majority of the gating current that precedes
the ion current in the Shaker K+ channel [6]. It is generally
believed that voltage dependent ion channels are gated by
translocation of S4 positively charged arginines across the
membrane electric field [7]. However, the pKa of titratable
groups in a protein is subjected to dramatic shifts that may
change their ionization state [8]. This ionization state depends
on local microenvironmental conditions such as surface
electric potential, solvent polarity and ionic strength; the pKa
of a basic residue would decrease when exposed to positive
electrostatic potentials or to less polar microenvironments.
Recently, the titratable nature of arginines on S4 in the
bacterial Na+ channel (NaChBac) was shown, suggesting that
S4 arginines are not permanently charged, and the changes in
their ionization state may play a central role in gating [9].
Schulte et al. reported regulation of channel gating by
changes in the pKa of a basic gating residue [10]. The authors
showed that in proton dependent Kir channels, a pH-sensing
lysine (Lys-80 in Kir1.1), located close to the intracellular side
of TM1, undergo a pKa shift of more than 3 pH units. A series
of point mutations revealed that the positive electrostatic
potentials from two spatially close arginines destabilize the
protonated state of Lys-80, which therefore changes its
ionization state around the physiological pH range.
Analogously, pKa shifts in the gating residues of voltage
dependent ion channels (R1-R4) may arise from strong
potentials, induced by a highly focused electric field in their
microenvironment [11, 12].
 T. Paldi: Salt-Bridge Switching
The theoretical basis for deprotonation of S4 arginines
cause by voltage-dependent pKa shifts is discussed herein. A
new helical screw-based model, which is compatible with
many experimental observations, shows how voltage-
dependent pKa shifts of arginines on S4 are involved with the
conformational changes in the VSM.
Results
An empirical model rationalizes arginine discharge
during NaChBac gating
The helical screw model [13] describes a screw-like
movement of S4, where, at each step, an emerging arginine
reaches the spatial position of the adjacent arginine three
residues apart, through a ‘canal’ formed by S1, S2 and S3 (the
so-called, ‘gating pore’). The movement of S4 along the
gating pore is mediated by the sequential formation and
impairment of salt bridges between positively charged
residues on S4, and negatively charged residues on S1-S3.
However, the way in which these electrostatic interactions
break and reform during S4 movement upon channel gating
remains unclear.
It was previously shown that the coupling between R1 on
S4 and E43 on S1, stabilizes the closed state of NaChBac
[14]. The pH-dependent coupling of R1 with E43 during
channel activation implies that protonation of R1 reinforces
its electrostatic interaction with E43, thereby stabilizing the
channel closed state, whereas its deprotonation is believed to
facilitate the mobilization of S4 during channel activation [9].
To examine this possibility, a model describing S4 movement
as a result of charge elimination of R1 and R2, rather than
charge separation from a negatively charged residue, is
suggested.
The model requires two helical screw steps for channel
activation [14], and impairment of eight ion pairs (two pairs
in each of the four VSMs in a tetrameric channel), each of
which hold S4 in the closed position. If the impairments occur
by deprotonation of eight arginines rather than separation of
charges, the channel open probability (po) may be described
according to:
8 linearly extrapolated to more negative voltages. Perpendicular dotted
(1) 𝑝𝑜(𝑤𝑡) = 𝑝dep
where po(wt) is the open probability of the wild-type channel,
and pdep is the average probability that a single arginine
residue will convert to its deprotonated form. Since
substitution of R1K hinders the coupling with E43 [14], this
substitution may account for the disruption of four out of
eight ion pairs in the tetrameric channel. An increase in the
channel open probability would thus be expected, since S4 is
now restrained by the remaining four ion pairs, according to:
8 1/2
4
(2) 𝑝o(R1K) = 𝑝dep = (𝑝dep )
3
Replacement of pdep8 by po(wt) in Eq. (2), according to Eq. (1),
gives:
1/2
(3) 𝑝𝑜(𝑅1𝐾) = 𝑝𝑜(𝑤𝑡)
Manipulation of the experimental data, obtained for the
normalized G-V curve of the unmodified NaChBac at pH 7.4
[9], according to Eq. (3), predicts the experimental left shift
of the R1K channel mutant curve (Fig. 1, WT1/2). Notably, the
model does not predict changes in the slope of the G-V curve
of R1K, which could be derived from interactions of the
arginine side chain with the membrane lipids [15]. As channel
gating involves many different types of conformational
changes, prediction of a voltage left shift upon R1K mutation
implies that charge elimination by arginine deprotonation
serves as a bottleneck in a more complex gating process.
Since pdep = po(wt)1/8, according to Eq. (1), the average pKa of
a single arginine equals 7.4, as is the pH of the extracellular
solution (pHo), at the corresponding voltage of the WT1/8
curve midpoint (V0.5(WT1/8) = -87 mV; Fig. 1, dashed curve).
Fig. 1 Channel gating is mediated by charge elimination. A fit to the
two-state Boltzmann function of normalized steady-state activation
curves of unmodified NaChBac (solid circles) and R1K mutant
(open circles) at pH 7.4 is presented. According to the ‘activation
through deprotonation’ model, the square root of the unmodified
NaChBac G-V curve (WT1/2; solid curve) and WT1/8 (dashed curve),
represents the po after elimination of the four charges, and the pdep,
respectively. The arrow indicates the point where the WT1/8 is
lines indicate the pdep midpoint, where the average pKa of a single
arginine is 7.4 (V0.5= -87 mV)
A ‘ratcheted capacitor’ element underlies a VSM
mechanistic model
Placing the principles underlying Eq. (1) within a structural-
functional context is of great importance to an understanding
of the voltage sensing mechanism of the VSM. The
mechanistic model describes structural changes in the VSM
that are related to its electrical traits, and are compatible with
intramolecular electrostatic interactions within the VSM, as
 T. Paldi: Salt-Bridge Switching
defined by the helical screw model [13, 16], and based on
accumulated experimental data, as follows:
External and internal water-filled crevices in the VSM
focus the electric field on a thin, low dielectric septum lying
between them [12, 17]. Double substitutions of charged
residues in the VSM of voltage-dependent sodium channels
[18] and voltage-dependent potassium channels [19-21],
which enable ion permeation through the VSM gating pore,
point to the importance of arginines on S4 and their
counterpart acidic residues in maintaining this low dielectric
septum intact, probably by stabilizing a constriction at the
gating pore. The constraint of the S4 position at rest [14] for
each of these double mutant sets, in which R1 and R2 line the
pathway connecting the internal and external crevices [22],
and the fact that an Hv1 proton channel truncated below R2
preserves its functionality [23], suggest that this constriction
in the gating pore is held by no more than two successive
arginines on S4 that may interact with two acidic residues
(such as E43 on S1 and D60 on S2, in NaChBac) at any given
conformation of the VSM [16] (Fig. 2). As is illustrated in
Fig. 2 (right), R4 interacts with D60 in the channel open state
[24], near the interface between the intracellular crevice of
the VSM and the low dielectric septum in the gating pore
above it [22]. A second ion pair between R3 and E43 delimits
this constriction in the gating pore from the extracellular side.
The thin low dielectric septum, lying between a deep
internal crevice (below D60) and a shallow external crevice
(above E43), fits the 3-7 Å thickness (Fig. 2, yellow boxed
region) that separates an internal cavity of 20-25 Å from an
external cavity of 3 Å, as suggested by Islas and Sigworth
[25], and is consistent with experimental results [12, 26]. The
outer and inner water-filled crevices of the VSM may be
envisioned as a two-plate capacitor, in which “water wire”
molecules are trapped inside its insulator (Fig. 3). Normally,
the septum of the VSM (referred to as the capacitor’s
“insulator”) does not conduct ions; therefore, a high electric
field (~108 V/m) induced by changes in the membrane
potential may polarize the water molecules within the septum,
due to their dipole moment. The polarized dipole moment
would induce an electric field opposite the external electric
field from the two water-filled crevices (referred to as the
capacitor’s “plates”). An arginine residue would tend to
release its proton when it is exposed to the positive
electrostatic potential of the dipole, by reducing the pKa of its
guanidine group. Conversely, the pKa would increase when
the arginine is exposed to the negative charge of the dipole.
Thus, polarization of the dipole by the electric field across the
septum, would lead to counter-pKa shifts of R4 and R3 at the
open state (Fig. 3), thereby facilitating the inward movement
of S4, by generating a new set of local energetic minima [27]
resulting from neutralization of R4, and subsequent
interaction between the positively charged R3, and D60 (Fig.
4
4, step 1 to step 2). This process is repeated so long as an
arginine on S4 can stabilize the E43 on S1, and is terminated
when the VSM adopts a stable closed conformation (Rest), at
which R1-R4 are paired with acidic residues in the VSM (Fig.
4, step 3).
At rest, R2 is deprotonated, due to the positive potential
at the septum boundary that stabilizes the unpaired D60. The
model allows for movement of charged arginines on S4, just
before their deprotonation. This movement is directed by the
electric field that falls across the septum. Membrane
depolarization reverses the polarity of the low dielectric
septum, and drives the outward movement of S4 (Fig. 4, steps
4-6).
Voltage-dependent pKa shifts initiate VSM activation
If protonation/deprotonation of arginines are the bottleneck
of the gating process, as noted above, the entire gating
process could be described by an Henderson-Hasselbalch
based equation that describes the effect of the pKa shift, or the
titration of arginines by [H+], on channel gating, as follows:
𝑛
(4) 𝑝𝑜 = (
1
)
1+10𝑝𝐾𝑎 −𝑝𝐻
where pKa represents the average pKa of a single S4 arginine,
pH represents the pH in the relevant VSM crevice, and n is
the number of deprotonated arginines that affect S4
movement. Upon hyperpolarization of the membrane
potential from the open state, the positive electrostatic
potential of a hypothetical dipole moment (μ) that is pointed
toward the inner side of the septum (near R4/D60) would
decrease the pKa of R4, according to:
𝜇𝐸𝑠𝑒𝑝
∆𝑝𝐾𝑎 = −0.43
𝑘𝑇
where Esep is the electric field across the septum, k is the
Boltzmann constant, and T is the absolute temperature. If the
distance between the charges of the dipole is equivalent to the
septum thickness, the equation is rewritten as:
(5) ∆𝑝𝐾𝑎 = −0.43
𝜀𝑉𝑚
𝑘𝑇
where ε is the positive charge of the dipole pointing toward
the arginine, and Vm is the membrane potential. Accordingly,
the negative electric potential at the septum’s external
boundary near R3/E43 would increase the pKa of the R3 side
chain (Fig. 3). The pKa at any point during gating is given by:
𝑝𝐾𝑎 = 𝑝𝐾𝑎1 + ∆𝑝𝐾𝑎
where pKa1 is the initial value before the shift (i.e., at 0 mV),
and ΔpKa is the change in the pKa. Replacing ΔpKa with the
expression in Eq. (5), and placing the expression obtained for
pKa in Eq. (4) at a given pHo, gives:
𝑛
1
(6) 𝑝𝑜 = ( 0.43𝜀 )
1+exp10(𝑝𝐾𝑎1 −𝑝𝐻𝑜 −𝑉𝑚 )
𝑘𝑇
Global fitting of the experimental G-V data sets of NaChBac
at both of the pHo values to Eq. (6), resulted in pKa1 = 4.6 ±
0.1, and 0.43ε/kT = 31.8 ± 1.3, for n = 8 (Fig. 5). Although
 T. Paldi: Salt-Bridge Switching

Fig. 2 Ribbon representations of the VSM in NaChBac. Three putative prevailing conformations of NaChBac are shown: Rest
(closed; left), Intermediate-closed (middle), and Open (right) states. The low dielectric septum (designated by the yellow box)
separates a shallow, extracellular crevice from a deep intracellular crevice [25]. Negatively charged residues (red) on S1-S3 and
R1-R4 (blue) on S4 are labeled and presented as sticks. The coordinates of NaChBac models [16] were kindly provided by Dr.
H.R. Guy

Fig. 3 The septum of the VSM functions as a capacitor's

insulator. An analogy is drawn between the 3D crystal
structure of KvAP VSM (right) [49] and a two-plate
capacitor (left).Two successive salt bridges, formed by
arginines (blue) and acidic residues (red), delimit the VSM
septum, which separates the extracellular and intracellular
water-filled crevices of the protein. Water molecules within
the septum change their orientation, due to interactions with
the electric field (E) across the septum. A positive charge of
the dipole that is pointed toward an arginine residue causes
a pKa downshift, followed by a proton release

Fig. 4 The ‘ratcheted capacitor’ model of S4 movement in

NaChBac. A schematic representation of S4 movement
(orange rod) relative to S1-S3 (green rod). Induced
polarization of the low dielectric septum (highlighted by the
yellow box) by the membrane’s electric field accounts for a
pKa shift of arginines at the low dielectric septum boundaries,
and their subsequent deprotonation/protonation (dashed
arrows). ‘Depol’ and ‘Repol’ denote depolarizing or
repolarizing potentials, respectively. Plus signs (blue)
represent R1-R4 on S4; minus signs (red) represent acidic
residues on S1-S3. The gray box designates the putative
lipid/water interface. Interacting residues are depicted
according to Shafrir et al. [16]

5
 T. Paldi: Salt-Bridge Switching
Fig. 5 Changes in NaChBac gating upon pHo transition. G-V
data points of NaChBac at pHo 7.4 and 6.0 are taken from [9].
Predicted G-V curves, according to Eq. (6), are presented as
smooth lines (fit coefficients ± standard deviation of the fit:
pKa1 = 4.6 ± 0.1; [0.43ε/kT] = 31.8 ± 1.3; n = 8)
the pKa1 value of the upper arginine might appear extremely
low, it is compatible with a reported pKa of arginine residues
inside the pore of the acetylcholine receptor [28]. The value
of pKa1 represents an upper limit of pKa shift, assuming that
protonation completely dominates gating. However, if
arginine protonation becomes less dominant during gating,
due to increased domination of other driving forces, smaller
pKa shift values are expected (see Discussion). The fitted
value for 0.43ε/kT suggests that a change of ~30 mV in the
membrane potential corresponds to a pKa shift of 1 unit, and
is consistent with reorientation of two hypothetical dipole
moments (or a dipole with two positive charges on one end
and two negative charges on the other), since ε is equivalent
to two elementary charges (~3x10-19 C).
Arginine deprotonation underlies gating-pore currents
observed upon a single mutation at S4
The passive conductance of protons or cations through
naturally occurring VSMs, namely the Hv1 proton channels,
or through modified VSMs in ion channels (the so-called
“gating-pore currents” or “omega currents”) [12, 20, 29-31],
indicates that the septum that separates the outer and the inner
water-filled crevices of the VSM is hydrated all along.
However, the VSM of ion channels is naturally impermeable
to ions and features a ‘blocked channel’ behavior. Ions leak
through the VSM of ion channels would lead to diseases such
as the hypokalemic periodic paralysis disease [32]. Molecular
dynamics simulation based on the crystal structure of voltage-
dependent potassium channels [33, 34] and on a structural
model of NaChBac [22], show that the salt bridge formed
between R4 on S4 and an acidic residue on S2 (corresponding
to the salt bridge formed by R4/D60 in NaChBac at the open
state; Fig. 2, right) interrupts the continuity of the water chain
6
along the VSM. This implies a central role for salt bridge
formation in blocking the ion path through the VSM.
While the notion that the septum of the VSM is virtually
maintained by two successive arginines, serves as a rationale
for gating pore currents upon double substitution of S4
arginines, in tandem [18, 19], gating pore currents that arise
upon only a single substitution [12, 31] remain unexplained.
Yet, the ratcheted capacitor model suggested herein shows
that the co-occurrence of a single arginine substitution and
deprotonation of the adjacent arginine, situated at the
septum’s boundaries, would render a continuous water chain
between the outer and the inner water-filled crevices, thus
enabling ion leaks through the VSM.
Generation of VSM gating pore currents upon a single
substitution at S4 fits well the results of histidine scan assays
performed in the Shaker K+ channel [12, 29, 30], as is
described below. In a VSM substituted at R1 (Fig. 6, upper
panel), deprotonation of R2 would impair the electrostatic
interactions between R2 and its negatively charged
counterpart at hyper-polarizing membrane potentials. The
impairment of two successive electrostatic interactions at the
septum boundaries would open an ion conducting pathway at
the resting state with a linear I(ω)-V curve pattern (Fig. 6,
upper panel), as has been shown for R1H substitution in the
Shaker K+ channel [12]. A different I(ω)-V pattern is
observed upon R2 (Fig. 6, middle panel) or R3 substitutions
(Fig. 6, bottom panel), as was reported for the Shaker K +
channel [30]. According to the model, the parabolic-shaped
curve reflects the transient inward passage of ions through the
gating pore, which peaks at intermediate conformational
states of the VSM, and is blocked either at the resting or the
open states.
Discussion
Extracellular acidification reduces the maximal conductance,
and shifts the voltage required for activation toward more
depolarizing potentials (right-shifted G-V) in many voltage
dependent ion channels [2]. Against the background of the
helical screw model, the left-shifted G-V curve of NaChBac
upon R1K substitution (Fig. 1) may reflect a facilitated
outward movement of S4 during channel activation. The
predicted left shift of R1K, given by Eq. (3), favors charge
diminution or elimination, due to the lower affinity of the
lysine's side chain toward protons compared to that of
arginine, over separation of fixed charges. Following this
rationale, recharging of the arginine by extracellular
acidification functions in opposition to this process, by
securing the electrostatic interaction with a negatively
charged residue (e.g., E43 on S2), and thus stabilizing S4 in
the retracted (closed) state, as indeed was shown
experimentally [9]. Since R2 and R3 occupy the position of
 T. Paldi: Salt-Bridge Switching
R1 at each helical screw step during activation (Figs. 2 and
4), they likely resemble R1 with regard to their charge
elimination.
The effect of voltage on arginine deprotonation is,
however, less straightforward. Although voltage-dependent
pKa shifts in ion channels were previously proposed [35, 36],
they were not assumed to constitute a substantial factor in the
VSM gating mechanism. Here, channel gating is adequately
predicted by the Henderson-Hasselbalch based model, which
describes a series of voltage-dependent pKa shifts. It is
therefore expected that, so long as arginine
protonation/deprotonation is a limiting factor in the VSM
activation process, the model would be capable of predicting
steady-state channel gating. However, divergence from the
model would be expected when the ionization of arginines
becomes less limiting in gating, as is shown, for example, in
NaChBac mutants that hamper channel activation (see E43C
or R3C NaChBac mutants in [9]). The model-fitted
coefficient value of 0.43ε/kT suggests that the gating process
may be mediated by orientation of a molecule or molecules
with a dipole moment, which harbors two charges at each
pole. This situation is reminiscent of the dipole with three
charges at each pole, suggested by the Hodgkin and Huxley
model [37].
7
Fig. 6 Deprotonation-assisted gating pore currents.
Upon membrane repolarization, preclusion of two
salt bridges at the septum's boundaries due to a point
mutation (X#) and deprotonation of the adjacent
arginine (dashed arrow) on S4 (orange rod), opens
an ion conductance pathway through the septum
(vertical solid arrow). A salt bridge formed in the
septum renders the gating pore impermeable to ions
(red X arrowhead). The pattern of the I(ω)-V (right
pane) is thus determined by the position of the point
mutation on S4, relative to the deprotonation site,
and is consistent with results of histidine scan
analyses in the Shaker K+ channel [12, 29, 30]
Furthermore, trapped water within the septum of the
VSM may form a single-file array that enables proton
conductance [38], such as occurs in unmodified and modified
VSMs (e.g. [4, 31]). Since water has a dipole moment, the
water hydrating the VSM septum may affect gating by
reorientation in response to an external electric field [39] and
change the chemo-electrical properties inside the protein.
Still, to ascertain the role of water in voltage-sensing, further
quantitative analysis is required.
Among the existing models that describe the VSM
mechanism, the helical screw model [13] is often claimed to
be the most compatible with experimental results (e.g., [24,
40]). Still, the basic helical screw model fails to explain two
major issues: (i) the relatively low temperature dependency
of gating, which is incompatible with breakage of several salt
bridges during S4 displacement, as has been postulated by
Green [41]; and (ii) the observed fast component of the gating
current, preceding the slower main gating current [42].
Arginine deprotonation, as is suggested here by the ratcheted
capacitor model, reconciles the helical screw model with
these two issues by suggesting that: (i) charge elimination
precludes electrostatic interactions, thereby reducing the high
negative change in enthalpy produced by salt bridge
formation, and: (ii) the initial arginine
 T. Paldi: Salt-Bridge Switching
deprotonation/protonation upon changes in membrane
polarization, which is independent of S4 movement (Fig 4,
step 1 and 4), and therefore exhibits faster kinetics than the
subsequent deprotonations, which depend on much slower
structural transitions within the protein.
The initial fast proton transfer suggests that more charge
can be transferred during smaller S4 movement compared to
the occurrence of tethered charges, thus reconciling a large
gating charge with a relatively small S4 movement. One
protonation event and three deprotonations over only two
helical screw steps (Fig. 4) imply that relocation of protons,
rather than of tethered charges, constitutes the gating current
as was suggested previously [43, 44], and is well-matched
with ~16 elementary charges measured for NaChBac [45]. Of
note is that two helical screw steps are equivalent to ~9 Å
along the S4 axis (assuming S4 is an α-helix) and are
compatible with an S4 trajectory of 10 ± 5 Å, measured in the
Shaker channel [46]. Also, an S4 trajectory of two helical
screw steps is consistent with the position of S4 at rest, which
is only two helical-screw steps away from the open state [14]
and is in agreement with a three-state kinetic model (i.e., rest,
intermediate-closed, and open states; Fig. 2) that claims to
account for most K+ channel behavior [47].
Conclusions
It is tempting to hypothesize that the evolution of VSMs into
proton channels [4, 5], or into proton sensors [48] is related
to the titratable nature of arginines on S4: that is to say, that
VSM modulation by protons originates from the function of
these proteins as voltage-to-proton transducers, as was
suggested by Green and colleagues [44]. The steep voltage
dependence, and the voltage threshold for channel activation
that thus far has been rationalized by empirical models (e.g.,
ref. 47), yields here a mechanistic explanation, in which a
voltage-dependent salt-bridge switching mechanism triggers
the helical-screw-like movement of S4.
Acknowledgments
The author thanks Dr. Michael E. Green for helpful
discussions, and for critical reading of the manuscript.
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