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Evaluation of synthetic linear motor-molecule actuation energetics

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Evaluation of synthetic linear motor-molecule actuation energetics
Evaluation of synthetic linear motor-molecule
actuation energetics
Branden Brough†‡, Brian H. Northrop§¶, Jacob J. Schmidt储, Hsian-Rong Tseng††, Kendall N. Houk§‡‡,
J. Fraser Stoddart§¶‡‡, and Chih-Ming Ho†‡,‡‡
Departments of †Mechanical and Aerospace Engineering, §Chemistry and Biochemistry, 储Bioengineering, and ††Molecular and Medical Pharmacology,
‡Institute for Cell Mimetic Space Exploration, and ¶California Nanosystems Institute, University of California, Los Angeles, CA 90095
By applying atomic force microscope (AFM)-based force spectroscopy together with computational modeling in the form of molecular force-field simulations, we have determined quantitatively
the actuation energetics of a synthetic motor-molecule. This multidisciplinary approach was performed on specifically designed,
bistable, redox-controllable [2]rotaxanes to probe the steric and
electrostatic interactions that dictate their mechanical switching at
the single-molecule level. The fusion of experimental force spectroscopy and theoretical computational modeling has revealed
that the repulsive electrostatic interaction, which is responsible for
the molecular actuation, is as high as 65 kcal䡠molⴚ1, a result that is
supported by ab initio calculations.
computational modeling 兩 force spectroscopy 兩 molecular motors 兩
switchable rotaxanes
M
olecular motors have recently garnered considerable interest within the domains of microsciences and nanosciences (1, 2). Harnessing the ability to selectively, cooperatively, and repeatedly induce structural changes in molecules
may hold the promise of engineered systems that operate with
the same complexity, elegance, and efficiency as biological
motors function in the human body. In natural systems, it has
become apparent that both macro and micro processes are
initiated and controlled by nanoscale molecular motors (3). For
example, myosin and kinesin are associated with muscle contraction and intracellular trafficking, respectively, and have
recently found their ways into engineered devices (4, 5). Moreover, initial work has been performed that demonstrates the
ability of natural nanoscale molecular motors to power microfabricated systems (6).
Synthetic motor-molecules (7–11), which are designed to excel
where their biological counterparts fall short, also have been
investigated. Whereas devices powered by biological molecules
require (4–6) chemical diffusion for actuation stimulus, synthetic molecules have been shown (2, 9) to operate with a variety
of different stimuli, thereby lending much greater flexibility to
a particular system’s design. Moreover, a synthetic nanoscale
actuating molecule carries with it an inherent ability to be
modified and optimized precisely for a specific task.
Switchable, bistable rotaxanes (2, 9), compounds comprised of
a dumbbell-shaped component containing two different recognition sites for an encircling ring-shaped component, show
particular promise as molecular actuators, given their ability to
undergo controllable, reversible mechanical switching with the
appropriate chemical, electrochemical, or photochemical stimulus in solution. Toward the goal of device applications, switching has been shown to operate in condensed phases such as in a
polymer electrolyte gel (12), on a self-assembled monolayer
(SAM) (13), on the solid supports of engineered systems (14),
and in molecular switch tunnel junctions (15). Bistable rotaxanes
benefit from their synthesis being highly modular, a virtue that
allows for a considerable degree of flexibility in their design.
Recently, the use of linearly actuating rotaxanes in microfabricated molecular devices has been demonstrated (16) where the
www.pnas.org兾cgi兾doi兾10.1073兾pnas.0509645103
redox-controlled switching of palindromic, doubly bistable [3]rotaxane molecules self-assembled on gold-coated microcantilevers was shown to generate enough surface stress to bend them
reversibly and repeatedly. The successful operation of these
fundamental devices demonstrates the potential for such rotaxanes to form the foundation on which a bottom-up fabricated
actuator paradigm based at the nanoscale could be built.
In addition to the actuation characteristics of synthetic
motor-molecules and their potential to be redesigned, the
possibility of producing forces greater than their natural
counterparts (16, 17) is a distinct benefit. Not only is this
assertion critical to the pursuit of artificial motor-molecule
engineering, but its verification is essential to any system’s
design. Force spectroscopy using an atomic force microscope
(AFM) has been demonstrated to be an effective probe (18) of
the strengths of various covalent bonds (19) as well as the
noncovalent bonding associated with the biotin–streptavidin
complex (20) and hydrophobic cyclodextrin-based complexes
in water (21). In addition, Gaub and coworkers (22) have used
AFM force spectroscopy to investigate the force generated by
polyazobenzenes operating as molecular motors.
Along that same vein, we have expanded this field of
research to include the probing of steric and electrostatic
interactions within highly complex synthetic molecules. In so
doing, we have been able to quantify the power stroke of the
switchable, bistable [2]rotaxane R4⫹ (Fig. 1), which can function as a linear motor-molecule by harnessing the redoxcontrolled mechanical shuttling of its ring along its dumbbell.
Although it features the same actuation mechanism as the
doubly bistable [3]rotaxane recently used to drive a set of
microcantilevers (16), R4⫹ was specifically designed (i) with a
thioctic acid tethered to its ring component for attachment to
a gold AFM tip and (ii) with a hydroxymethyl group on one of
the stoppers of its dumbbell for attachment (Scheme 1b) to
SiO2 via covalently bound monolayers of isocyanatopropyl
linkers (23). This dual functionality allows for AFM force
spectroscopy studies to probe the steric and electrostatic
interactions present in the ground (Fig. 1a) and oxidized (Fig.
1b) states of R4⫹. AFM probing of molecules in neat ethanol
(EtOH) will measure the repulsive steric interactions (Fig. 2a)
between the cyclobis(paraquat-p-phenylene) (CBPQT4⫹) ring
and the diisopropylphenyl ether stopper and serves as a
control. The probing of the R6⫹ molecules in an oxidizing
solution will measure the repulsive force (Fig. 2b) between the
CBPQT4⫹ ring and an oxidized tetrathiafulvalene (TTF2⫹),
which is responsible for molecular actuation. In conjunction
Conflict of interest statement: No conflicts declared.
This paper was submitted directly (Track II) to the PNAS office.
Abbreviations: AFM, atomic force microscope兾microscopy; CBPQT4⫹, cyclobis(paraquat-pphenylene); TTF, tetrathiafulvalene; DNP, 1,5-dioxynaphthalene; SAM, self-assembled
monolayer.
‡‡To
whom correspondence may be addressed. E-mail: [email protected], [email protected]
chem.ucla.edu, or [email protected]
© 2006 by The National Academy of Sciences of the USA
PNAS 兩 June 6, 2006 兩 vol. 103 兩 no. 23 兩 8583– 8588
APPLIED PHYSICAL
SCIENCES
Edited by Julius Rebek, Jr., The Scripps Research Institute, La Jolla, CA, and approved April 21, 2006 (received for review November 14, 2005)
Fig. 1. Structural formula and schematic representation of bistable [2]rotaxane R4⫹, in which an electron-poor cyclobis(paraquat-p-phenylene) (CBPQT4⫹) ring is confined to a dumbbell containing two electron-rich recognition sites, tetrathiafulvalene (TTF) and 1,5-dioxynaphthalene (DNP), by the
presence of bulky 2,6-diisopropylphenyl ether stoppers at each end. One of
these stoppers, the one closer to DNP, carries a hydroxymethyl group on its
4-position for subsequent attachment to silicon wafers. (a) The CBPQT4⫹ ring,
which carries a tether terminated by a thioctic acid ester for attachment to a
gold-coated AFM tip, displays a stronger interaction with TTF than with DNP
and thus resides selectively (16) on the former. (b) Chemical oxidation of TTF
to TTF2⫹ results in a strong charge– charge repulsion between the CBPQT4⫹
ring and TTF2⫹, a situation that causes the CBPQT4⫹ ring to shuttle to DNP (9,
12–16) in the oxidized [2]rotaxane R6⫹. The mechanical movement of the
CBPQT4⫹ ring from TTF2⫹ to DNP resembles the power stroke of a linear motor.
Reduction of the TTF2⫹ to its neutral state (TTF) prompts the ring to shuttle
back thermally from a metastable state to its ground state where it encircles
TTF in a manner reminiscent of the diffusive stroke of a linear motor. This
externally controllable shuttling process and bistability allows [2]rotaxane R4⫹
to function as a nanoscale linear motor given its ability to contract and expand
reversibly in the presence of a chemical oxidant or reductant, respectively.
with computational chemistry, these experiments enable a
thorough quantitative evaluation of the actuating energetics of
a single-molecule motor. For further details, see Supporting
Materials and Methods, Tables 1 and 2, Figs. 5–16, and Scheme
2, which are published as supporting information on the PNAS
web site.
Results and Discussion
The routes used to synthesize the dumbbell-shaped compound
3 and the bifunctional [2]rotaxane R4⫹ are shown in Scheme
1a, and that used to covalently attach [2]rotaxane R4⫹ to SiO2
wafers is shown in Scheme 1b. The concentration of R4⫹ was
kept dilute, ⬇1 per 100 nm2, to promote the observation of
single-molecule events. Compiled AFM force probe data are
summarized (Fig. 3) in the form of histograms, which were
analyzed by using commercial peak-finding and curve-fitting
algorithms. The first histogram (Fig. 3a) summarizes the
control experiments on unoxidized single molecules and reveals that the most probable molecular rupture force occurs at
74 pN. In these control experiments, any measured force peak
stems from rupture either within the molecule or its linkages
to the AFM tip or the substrate. As in any structural system,
failure will occur at the weakest point. Published reports
indicate that this ruptured 74-nN mechanical barrier is considerably weaker than any of the covalent bonds in the system,
revealing that the failure occurs as the ring passes over the
stopper, a process referred to as ‘‘deslipping’’ (24). To our
knowledge, all experimental and theoretical data at comparable experimental conditions indicate that the rupture of any
covalent bond requires ⬎1 nN (see Table 2).
The second histogram (Fig. 3b) summarizes the results of
force probing in the presence of a 10⫺4 M Fe(ClO4)3 oxidizing
enthanolic solution, which has been shown to induce ring
translation to the 1,5-dioxynaphthalene (DNP) site as a result
of electrostatic repulsion from the TTF2⫹ site (14). Whereas
in the unoxidized situation the histogram reveals a singular
force, this histogram indicates that there are two distinct force
barriers, one at 66 pN (O1, solid blue curve) and the other at
145 pN (O2, solid green curve). We believe that the lower force
regime is caused by the probing of single molecules that have
not undergone redox-controlled switching despite the oxidizing environment. This belief is supported by comparing the
peak value from the unoxidized histogram (U, dashed blue
curve) with those from the oxidized histogram (Fig. 3b). The
peak of the curve for the unoxidized molecules coincides well
with the lower force peak curve from the oxidation experiments. This interpretation also is supported by earlier studies
Scheme 1. Reaction schemes illustrating the synthesis of dual recognition [2]rotaxane R4⫹ (a) and the attachment of R4⫹ to SiO2 wafers via covalently bound
isocyanatopropyl tethers (b).
8584 兩 www.pnas.org兾cgi兾doi兾10.1073兾pnas.0509645103
Brough et al.
at a lower concentration of oxidant (10⫺6 M), which yielded
results that were not easily distinguishable from those obtained
for the unoxidized molecule with respect to peak-force magnitude and well defined peak distribution (see Fig. 16). An
alternative explanation could be that electron transfer between the Fe(ClO4)3 oxidant and TTF occurs within the time
frame of individual pulling experiments, resulting in an equilibrium of neutral and oxidized TTF, which would give rise to
both unoxidized as well as oxidized force data. We believe,
however, that results of probing an oxidant concentration of
10⫺6 M support the former explanation over the latter.
Although the second force regime at 145 pN has clearly
emerged as a result of the presence of oxidant, it is more
important to examine how these results enhance our understanding of the bistable rotaxane’s mechanical switching behavior.
Previous experimental results have revealed that Fe(ClO4)3
oxidizes the TTF unit to TTF2⫹, resulting in concomitant
Brough et al.
Fig. 3. Force probe data obtained during single-molecule investigations of
R4⫹ and R6⫹. (a) Histogram of compiled AFM force spectroscopy data of
ground-state [2]rotaxane R4⫹ obtained in an EtOH solution with a loading rate
of 6 pN䡠s⫺1. The dashed blue curve (U) is obtained from commercial peak
finding and curve-fitting algorithms and indicates a most probable rupture
force of 74 pN. (b) Histogram of AFM force spectroscopy data obtained (for
R6⫹) in an oxidizing solution of 10⫺4 M Fe(ClO4)3 in EtOH (loading rate of 6
pN䡠s⫺1) displays two different force regimes, indicating two distinct force
barriers having most probable rupture forces of 66 pN (O1, solid blue curve)
and 145 pN (O2, solid green curve). The magnitude of the solid blue curve
matches well with the dashed blue curve superimposed from a, indicating that
not all molecules are oxidized. The solid green curve corresponds to the
emergence of a new, larger barrier, which is a direct measure of the electrostatic repulsion between the TTF2⫹ dication and the CBPQT4⫹ tetracation
responsible for molecular actuation.
shuttling of the CBPQT4⫹ ring to the DNP unit (9, 12–16) even
when immobilized on a surface or between surfaces in a manner
not dissimilar to that present in the AFM-based system. Therefore, the new higher-energy barrier can be ascribed to the
electrostatic repulsion between TTF2⫹ and CBPQT4⫹ or covalent bond rupture. Given that the measured peak rupture force
in the presence of the oxidant is still an order of magnitude lower
than those associated with the rupture of relevant covalent bonds
(see Table 2), it is clear that the second force regime present in
the experiments on the oxidized molecules emerges as a result of
the newly generated repulsive electrostatic barrier, which is
ultimately responsible for the molecule’s actuation.
Although the results indicate that the 145-pN force peak is a
measurement of the electrostatic repulsion between the bistable
rotaxane’s CBPQT4⫹ ring at TTF2⫹ dication, and therefore a
good indication of the maximum switching force producible
under load by the oxidized molecule, force spectroscopy meaPNAS 兩 June 6, 2006 兩 vol. 103 兩 no. 23 兩 8585
APPLIED PHYSICAL
SCIENCES
Fig. 2. Schematics of AFM force spectroscopy of [2]rotaxane R4⫹. (a) Groundstate probing in the absence of oxidant. (i) [2]Rotaxane R4⫹ confined to a
silicon surface and attached to a gold AFM tip via two gold–sulfur bonds. (ii)
Retraction of the AFM tip from the surface pulls the CBPQT4⫹ ring away from
the TTF unit and toward the bulky stopper. (iii) Continued retraction causes
the CBPQT4⫹ ring to pass over the stopper (deslip) at which point the force
required to overcome this physical barrier is measured. (b) Oxidized-state
probing performed in the presence of 10⫺4 M Fe(ClO4)3 in EtOH. (iv) The
attachment of a gold AFM tip to [2]rotaxane R6⫹ whose ring is now located on
the lower DNP unit as a result of the chemically induced oxidation of the TTF
unit to TTF2⫹ and subsequent electrostatic repulsion of CBPQT4⫹. (v) Retraction of the AFM tip forces the tetracationic CBPQT4⫹ ring toward the dicationic
TTF2⫹ despite the electrostatic repulsion between the two moieties. The AFM
will measure the repulsive force between the ring and the TTF2⫹ that was
originally responsible for the molecule’s actuation. (vi) Continued retraction
of the AFM tip enables the CBPQT4⫹ ring to overcome the electrostatic barrier
imposed by TTF2⫹, resulting in rupture of the mechanical bond posed by the
stopper. When measuring the higher-energy electrostatic repulsion of the
oxidized molecule, the subsequent deslipping event will not be measured
because of the dynamic instability of the AFM probe.
first-order chemical process to the activation energy, Ea, associated with that process
kr ⫽ Ae
⫺
冉 冊⫽
Ea
RT
1
,
t off
where A is a constant preexponential factor and R is the molar
gas constant. By substituting this relationship into Evans’ equation, a new relationship arises that describes the energy difference between two rupture events at a single loading rate
E a 1 ⫺ E a2 ⫽
Fig. 4. The ground-state energy profile obtained from molecular dynamics
and mechanics simulations. The x axis indicates the position of the CBPQT4⫹
ring along the linear dumbbell (in Å), and the y axis displays relative energies
of the different CBPQT4⫹ ring co-conformers (in units of kcal䡠mol⫺1). The
simulations correctly predict the co-conformation of the CBPQT4⫹ ring stationed on TTF to be the lowest-energy state and therefore the most favorable
one. The energy barrier that exists when the CBPQT4⫹ ring passes over either
of the bulky stoppering units (deslipping) is calculated to be 46 kcal䡠mol⫺1. A
combination of theoretical modeling and experimental AFM measurements
predict oxidation of [2]rotaxane R4⫹ to result in a 65-kcal䡠mol⫺1 repulsion
between the CBPQT4⫹ tetracation and the TTF2⫹ dication. This result is supported by quantum mechanical calculations. The dashed green line is a
schematic representation of what the full, oxidized energy profile may look
like, but it is not quantitative.
surements require loading-rate analysis for a complete understanding of the energy barrier that was investigated. A direct
corollary between force and energy through distance cannot be
established because the work done by an AFM’s tip distorts the
system’s energy landscape. According to Evans (25), forces
measured by force spectroscopy will be altered by thermal
fluctuations as a function of the probe’s loading rate such that
the most probable measured force, f*, is defined as
f* ⫽
冋
冉 冊册
t off x ␤
k BT
ln共k s␯ 兲 ⫹ ln
x␤
k BT
,
[1]
where kBT is the thermal energy, x␤ is the bond length, toff is the
lifetime of the bond under no external force, and ks and ␯ are the
spring constant and retract velocity of the AFM probe, respectively. Previously, only with a detailed loading-rate analysis that
spans several orders of magnitude can interactions be described
(20, 25) by x␤ and toff. Although rotaxanes do operate reversibly
(2, 9, 12–16), the irreversible rupture of molecules required by
this experiment (see Loading Rate Analysis in Dynamic Force
Spectroscopy in Supporting Materials and Methods) prohibits such
a study. However, by augmenting the AFM data with molecular
dynamics simulations (see Fig. 10), which describe (Fig. 4) the
ground-state energy profile of R4⫹ as a function of the ring’s
position on the dumbbell, an accurate description of the physical
switching mechanism can be formulated.
toff is the time at which a bond, described by an energy barrier
with Arrhenius dependence, spontaneously ruptures (25, 26).
Conversely, the Arrhenius equation relates the rate, kr, of a
8586 兩 www.pnas.org兾cgi兾doi兾10.1073兾pnas.0509645103
R
kB
冋
f 1*x ␤1 ⫺ f 2* x ␤2 ⫺ k BT ln
x ␤1
x ␤2
册
.
By using the measured force spectroscopy values of 66 pN for the
deslipping barrier under nonoxidative conditions and 145 pN for
the oxidant-induced electrostatic repulsion barrier, along with x␤
values, produced from molecular simulations, of 8.6 Å for the
deslipping of the CBPQT4⫹ ring over the bulky stopper and 13.0
Å as the distance necessary for the CBPQT4⫹ ring to travel from
the DNP recognition site to the TTF2⫹ dication, the difference
in interaction energies can be calculated as 19 kcal䡠mol⫺1.
Although the value of 66 pN was generated in the presence of
oxidant, this value was used instead of the 74-pN value produced
in the nonoxidizing environment because of the availability of
significantly more experimental data points. It should be noted
that the use of the 74-pN value only changes the results of this
study by 1 kcal䡠mol⫺1. By combining the 19 kcal䡠mol⫺1 difference
in oxidative and nonoxidative interaction energies with the
theoretically determined value of 46 kcal䡠mol⫺1 for the energy
associated with ground-state deslipping (Fig. 4), a final value of
65 kcal䡠mol⫺1 was obtained, representing the amount of repulsive actuation energy between CBPQT4⫹ and TTF2⫹ produced
by the oxidation of R4⫹ (Fig. 4). As measured by this approach,
65 kcal䡠mol⫺1 represents the upper limit of the total actuation
energy. In the experimental set-up, the retraction of the AFM tip
forces the CBPQT4⫹ ring into the most energetically unfavorable
conformation possible with respect to the TTF2⫹, thus maximizing the electrostatic repulsion between the tetracationic and
dicationic moieties.
This value, although seemingly low for the repulsion of six
positive charges confined within a radius of ⬇5 Å, is strongly
supported by ab initio calculations (see Model System and
Calculations of Electrostatic Repulsion in Supporting Materials
and Methods). Single-point quantum mechanical calculations
were performed in EtOH by using atomic coordinates from the
crystal structures of TTF, [TTF][ClO4]2, [CBPQT][PF6]4, and
[CBPQT傺TTF][PF6]4 as well as on five model systems for the
hexacationic [CBPQT傺TTF][Cl]6 complex (see Fig. 11). The
repulsive energy within this hexacationic complex was calculated
to be 70.6–76.0 kcal䡠mol⫺1. By using this value and that produced
by the molecular force field simulations, computational analysis
predicts that the electrostatic barrier resulting from the TTF2⫹
dication exceeds the steric barrier of the stopper by 25–30
kcal䡠mol⫺1. This entirely theoretical value compares well with
the value of 19 kcal䡠mol⫺1 produced experimentally by means of
AFM force spectroscopy. The similarity between the purely
independent ab initio calculations, which only take into account
the interaction between CBPQT4⫹ and TTF2⫹, and experimental AFM results lends support to the conclusion that singlemolecule rupture events are being measured accurately as 65
kcal䡠mol⫺1. Perhaps even more significant is the fact that this
result lends validity to the methods developed in this work used
to analyze force spectroscopy experiments at a single loading
rate with the help of molecular force-field simulations.
Employing a combination of AFM force spectroscopy and
molecular simulations, we have determined and verified that the
Brough et al.
Materials and Methods
General Methods. Chemicals were purchased from Aldrich and
used as received. Silicon wafers were purchased from Silicon
Quest International (Santa Clara, CA). The TTF-containing
tosylate 1 (27), the DNP-containing diol 2 (28), and the dication
4䡠2PF6 (27) were all prepared according to procedures described
in the literature. Solvents were dried following methods described in the literature (29). All reactions were carried out
under an anhydrous argon兾nitrogen atmosphere. Thin-layer
chromatography (TLC) was performed on aluminum sheets
coated with silica-gel 60F (Merck 5554). The plates were inspected by UV light and, if necessary, developed in I2 vapor.
Column chromatography was carried out by using silica-gel 60
(Merck 9385; 230–400 mesh). Melting points were determined
on a melting point apparatus (Model 9100, Electrothermal,
Dubuque, IA) and are uncorrected. All 1H and 13C NMR spectra
were recorded on either (i) an ARX500 (500 and 125 MHz,
respectively) or (ii) an Avance500 (500 and 125 MHz, respectively) (both from Bruker, Billerica, MA) by using residual
solvent as the internal standard. Samples were prepared by using
CD3COCD3 or CD3CN purchased from Cambridge Isotope
Laboratories (Cambridge, MA). All chemical shifts are quoted
by using the ␦ scale, and all coupling constants are expressed in
hertz. Matrix-assisted laser desorption ionization spectra
(MALDI) were recorded on a instrument from PerSeptive
Biosystems (Framingham, MA). Electrospray mass spectra
(ESI) were measured on a ProSpec triple focusing mass spectrometer (VG Analytical, Manchester, U.K.).
linked to the tethers upon reaction of its terminal alcohol with
the isocyanate of the SAM. Unreacted isocyanates were capped
with MeOH. Surface coverage and quality were evaluated
through hydrophilicity and homogeneity studies. Freshly prepared SAMs were stored under Ar(g) in glass vials until use.
Force Spectroscopy. AFM-based dynamic force spectroscopy stud-
ies were performed at standard temperature and pressure with
a Multimode scanning probe microscope with a Nanoscope 3A
controller and Pico Force system (Digital Instruments兾Veeco
Probes, Santa Barbara, CA), both in degassed 200 proof EtOH
and in the presence of an oxidizing solution of 10⫺4 M Fe(ClO4)3
in EtOH, which is consistent with previous work that demonstrated mechanical molecular switching within highly packed
rotaxane-based thin films (14). Samples were exposed to the
oxidant for no less than 10 min to ensure oxidation of the surface
bound molecules but no more than 90 min to guarantee the
integrity of the SAM. Control experiments for ground-state
molecule investigation were conducted in the presence of pure
EtOH. The nonaqueous solvent allowed the molecules to be
exposed to a plasma-cleaned, Au-coated AFM tip for thiol bond
formation. The bio-lever probe tip (Asylum Research, Santa
Barbara, CA) was lowered to the surface and held at that
position for 3 s at a force of 1.0–1.2 nN and then raised at a rate
of 1.05 ␮m䡠s⫺1. This procedure resulted in an ⬇10% probability
of specific bond formation, thereby minimizing the probability of
multiple molecule binding events. Specific bond formation was
assumed to have taken place when a measurable rupture force
occurred while the tip was no longer in contact with the
substrate. Each AFM probe had an average spring constant of 6
pN䡠nm⫺1 leading to a loading rate of ⬇6 nN䡠s⫺1. Individual
spring constants were measured by using the on-board thermaltuning program, and data points were analyzed with their
associated probes’ spring constant values. Experiments were
conducted with new samples, fresh solution, and new probes and
continued until the tip surface was saturated, as indicated by the
lack of any binding events despite a change in probe position.
Data were plotted in histograms prepared by using ORIGINPRO
software (OriginLab, Northampton, MA). Different bin sizes did
not affect the analysis or conclusions drawn from the data. Peaks
were identified and analyzed by using the Lorentzian function
that best fit our data. In the case of the multiple peak data, the
Levenberg–Marquardt algorithm was used in association with
the Lorentzian function.
Molecular Force-Field Simulations of R4ⴙ. A ground-state energy
Synthesis of Dumbbell-Shaped Compound 3 and [2]Rotaxane R䡠4PF6. A
MeCN solution of the tosylate 1 (27), the diol 2 (28), K2CO3,
LiBr, and 18C6 was heated under reflux for 16 h. After work-up,
the crude product was subjected to column chromatography
(SiO2:EtOAc兾hexane, 1:1) to give 3 as a yellow solid in 82%
yield.
The synthesis of R4⫹ was completed by stirring a solution of
3, 1,4-bis(bromomethyl)benzene, and the dication 4䡠2PF6 (27) in
anhydrous dimethylformamide (DMF) at room temperature for
10 d. The reaction mixture then was subjected directly to column
chromatography (SiO2), and unreacted 3 was recovered with
Me2CO, whereupon the eluent was changed to Me2CO:NH4PF6
(1.0 g of NH4PF6 in 100 ml of Me2CO), and the green band
containing the [2]rotaxane R䡠4PF6 was collected. After removal
of solvent, H2O was added, and the resulting precipitate was
collected by filtration to afford 57% yield of [2]rotaxane R䡠4PF6
as a green solid.
Attachment of R4ⴙ to SiO2. A SAM of covalently bound isocyana-
topropyl tethers (23) was formed by vapor deposition of 3(triethoxysilyl)propyl isocyanate in PhMe onto BOE and
piranha-treated 1 cm2 SiO2 wafers. R4⫹ then was covalently
Brough et al.
profile representing the relative energies of the different coconformations of R4⫹ as the CBPQT4⫹ ring is moved along the
dumbbell was generated by using the program MAESTRO V3.0.038
(30) with the AMBER* force field (31) and GB兾SA solvent
model (32) for CHCl3. Electrostatic point charges were calculated at the HF兾6-31G* level by using the program GAUSSIAN 03
(33) and fit to each atom. To model the ring’s movement along
the dumbbell, a ‘‘dummy’’ atom was placed at a fixed distance
from the model system and constraints were used to ‘‘pull’’ the
CBPQT4⫹ ring along the linear dumbbell in specified increments
(see Fig. 10). A 50-ps molecular-dynamics simulation (1.5-fs time
step) at a simulation temperature of 500 K, followed by energy
minimization of 200 randomly selected co-conformations, was
used for co-conformational searching at each fixed distance
along the dumbbell reaction coordinate. The output of each step
then was used as the input for the next. Energies were normalized relative to the overall lower-energy co-conformer of R4⫹,
which corresponds to the CBPQT4⫹ ring encircling the TTF unit.
The resulting energy differences (⌬E) plotted against CBPQT4⫹
ring position gave the ground-state energy profile of R4⫹. The
same method also was applied to a series of single-station
[2]rotaxanes containing stoppers of different sizes (see Fig. 8) as
PNAS 兩 June 6, 2006 兩 vol. 103 兩 no. 23 兩 8587
APPLIED PHYSICAL
SCIENCES
energy output of a single synthetic motor-molecule operating
within an engineered environment is 65 kcal䡠mol⫺1. Assuming
that the total energy available to biological systems from the
hydrolysis of ATP (14.4 kcal䡠mol⫺1 at physiological concentrations) (1) is 100% efficiently converted, rotaxanes still outperform nature by ⬎4.5 times. Furthermore, the synthetic origin of
switchable, bistable rotaxanes allows them to be designed and
redesigned according to a particular application’s requirements
as well as from the feedback obtained through characterization
studies. Molecular-level characterization is a critical step in the
iterative process that may eventually bring about viable engineered systems that extend far beyond mere scientific novelties.
Meanwhile, the promise of up to 65 kcal䡠mol⫺1 of energy
production from bistable rotaxanes and its family of synthetic
molecules will always headline systems that have the potential to
exceed anything seen in nature.
well as a degenerate [2]rotaxane containing two DNP recognition units and compared with experimental results to access the
viability of the molecular modeling procedure. In the case of the
degenerate [2]rotaxane, modeling predicted a shuttling barrier
of 19 kcal䡠mol⫺1, which is within 3 kcal䡠mol⫺1 of experimental
value of 16 kcal䡠mol⫺1 (34).
Model System and Calculations of Electrostatic Repulsion Between
CBPQT4ⴙ and TTF2ⴙ. Quantum mechanical calculations were used
to theoretically examine the electrostatic repulsion between the
CBPQT4⫹ tetracation and the TTF2⫹ dication that is responsible
for redox-controlled mechanical actuation of R4⫹. Single-point
quantum-mechanical calculations were performed at the HF兾
6-31⫹G* level by using the program GAUSSIAN 03 (33). Geometries of TTF (35), [TTF][ClO4]2 (36), [CBPQT][PF6]4 (37), and
the [CBPQT傺TTF][PF6]4 complex (38) were taken from x-ray
crystal data (see Fig. 11). To minimize computational cost,
all counterions present in crystal structures of charged species
were replaced with Cl⫺ anions. As a result of its energetic
instability, no cr ystal data exists for the hexacationic
[CBPQT傺TTF][PF6]6 complex. Therefore, five model systems
1. Howard, J. (2001) Mechanics of Motor Proteins and the Cytoskeleton (Sinauer,
Sunderland, MA).
2. Chatterjee, M. N., Kay, E. R. & Leigh, D. A. (2006) J. Am. Chem. Soc. 128,
4058–4073.
3. Kreis, T. & Vale, R. (1999) Guidebook to the Cytoskeletal and Motor Proteins
(Oxford Press, Oxford), 2nd Ed.
4. Limberis, L. & Stewart, R. J. (2000) Nanotechnology 11, 47–51.
5. Xi, J. Z., Schmidt, J. J. & Montemagno, C. D. (2005) Nat. Mater. 4, 180–184.
6. Soong, R. K., Bachand, G. D., Neves, H. P., Olkhovets, A. G., Craighead, H. G.
& Montemagno, C. D. (2000) Science 290, 1555–1558.
7. Kelly, T. R. (2001) Acc. Chem. Res. 34, 514–522.
8. Koumura, N., Geertsema, E. M., van Gelder, M. B., Meetsma, A. & Feringa,
B. L. (2002) J. Am. Chem. Soc. 124, 5037–5051.
9. Balzani, V., Credi, A., Raymo, F. M. & Stoddart, J. F. (2000) Angew. Chem. Int.
Ed. 39, 3349–3391.
10. Hawthorne, M. F., Zink, J. I., Skelton, J. M., Bayer, M. J., Liu, C., Livshits, E.,
Baer, R. & Neuhauser, D. (2004) Science 303, 1849–1851.
11. Badjić, J. D., Balzani, V., Credi, A. & Stoddart, J. F. (2004) Science 303,
1845–1849.
12. Flood, A. H., Peters A. J., Vignon, S. A., Steuerman, D. W., Tseng, H. R., Kang,
S., Heath, J. R. & Stoddart, J. F. (2004) Chem. Eur. J. 10, 6558–6564.
13. Tseng, H.-R., Wu, D., Fang, N., Zhang, X. & Stoddart, J. F. (2004)
ChemPhysChem. 5, 111–116.
14. Huang, T. J., Tseng, H.-R., Sha, L., Lu, W. X., Brough, B., Flood, A. H., Yu,
B. D., Celestre, P. C., Chang, J. P., Stoddart, J. F., et al. (2004) Nano. Lett. 4,
2065–2071.
15. Flood, A. H., Stoddart, J. F., Steuerman, D. W. & Heath, J. R. (2004) Science
306, 2055–2056.
16. Huang, T. J., Brough, B., Ho, C.-M., Liu, Y., Flood, A. H., Bonvallet, P. A.,
Tseng, H. R., Stoddart, J. F., Baller, M. & Magonov, S. (2004) App. Phys. Lett.
85, 5391–5393.
17. Fisher, M. E. & Kolomeisky, A. B. (1999) Physica A 274, 241–266.
18. Beyer, M. K. & Clausen-Schaumann, H. (2005) Chem. Rev. 105, 2921–2948.
19. Grandbois, M., Beyer, M., Rief, M., Clausen-Schaumann, H. & Gaub, H. E.
(1999) Science 283, 1727–1730.
20. Schmidt, J. J., Jiang, X. & Montemagno, C. D. (2002) Nano. Lett. 2, 1229–1233.
21. Auletta, T., de Jong, M. R., Mulder, A., van Veggel, F. C. J. M., Huskens, J.,
Reinhoudt, D. N., Zou, S., Zapotoczny, S., Schonherr, H., Vancso, G. J., et al.
(2004) J. Am. Chem. Soc. 126, 1577–1584.
8588 兩 www.pnas.org兾cgi兾doi兾10.1073兾pnas.0509645103
for the hexacationic [CBPQT傺TTF][Cl]6 complex were constructed by using the crystal structure of [CBPQT傺TTF][PF6]4
⫺
(38), with PF⫺
6 counterions replaced by Cl and two additional
counterions to balance the charge of oxidized TTF. The two
additional counterions were positioned within the most electropositive regions of the complex as determined by electrostatic
potential calculations (see Fig. 12 and 13). The five model
systems differed slightly in their positioning of the two new
counterions to produce a range of values for the repulsion
between [CBPQT][Cl]4 and [TTF][Cl]2. The CPCM solvent
model for EtOH was included to account for solvent effects.
We thank Tony Jun Huang, Paul A. Bonvallet, Amar H. Flood, Miguel
Garcia-Garibay, and Dean Astumian for valuable discussions and technical assistance. This work was supported in part by National Science
Foundation (NSF) Nanoscale Interdisciplinary Research Teams Grant
ECS-0103559, NSF Integrative Graduate Education and Research
Traineeship (Materials Creation Training Program) Fellowship DGE0114443 (to B.H.N.), and by the Defense Advanced Research Projects
Agency (DARPA) Biomolecular Motors program. Some of the compound characterizations are supported by NSF Equipment Grants
CHE-9974928 and CHE-0092036.
22. Hugel, T., Holland, N. B., Cattani, A., Moroder, L., Seitz, M. & Gaub, H. E.
(2002) Science 296, 1103–1106.
23. Haller, I. (1978) J. Am. Chem. Soc. 100, 8050–8055.
24. Asakawa, M., Ashton, P. R., Ballardini, R., Balzani, V., Belohradsky, M.,
Gandolfi, M. T., Kocian, O., Prodi, L., Raymo, F. M., Stoddart, J. F., et al.
(1997) J. Am. Chem. Soc. 119, 302–310.
25. Evans, E. (2001) Annu. Rev. Biophys. Biomol. Struct. 30, 105–128.
26. Bell, G. I. (1978) Science 200, 618–627.
27. Liu, Y., Flood, A. H., Bonvallet, P. A., Vignon, S. A., Northrop, B. H., Tseng,
H.-R., Jeppesen, J. O., Huang, T. J., Brough, B., Baller, M., et al. (2005) J. Am.
Chem. Soc. 127, 9745–9759.
28. Steuerman, D. W., Tseng, H.-R., Peters, A. J., Flood, A. H., Jeppesen, J. O.,
Nielsen, K. A., Stoddart, J. F. & Heath, J. R. (2004) Angew. Chem. Int. Ed. 43,
6486–6491.
29. Perrin, D. D. & Armarego, W. L. F. (1998) Purification of Laboratory Chemicals
(Pergamon, New York).
30. Mohamadi, F., Richards, N. G. J., Guida, W. C., Liskamp, R., Lipton, M.,
Caufield, C., Chang, G., Hendrickson, T. & Still, W. C. (1990) J. Comput. Chem.
11, 440–467.
31. Weiner, P. S. J., Kollman, P. A., Case, D. A., Singh, V. C., Ghio, C., Alagona,
G., Profeta, S., Jr., & Weinre, P. (1986) J. Am. Chem. Soc. 106, 765–784.
32. Still, W. C., Tempczyk, A., Hawler, R. C. & Hendrickson, T. (1990) J. Am.
Chem. Soc. 112, 6127–6129.
33. Frisch, M. J., Trucks, G. W., Schlegel, H. B. Scuseria, G. E., Robb, M. A.,
Cheeseman, J. R., Montgomery, J. A., Jr., Vreven, T., Kudin, K. N., Burant,
J. C., et al. (2004) GAUSSIAN 03 (Gaussian, Wallingford, CT), Revision C.02.
34. Kang, S., Vignon, S. A., Tseng, H.-R. & Stoddart, J. F. (2004) Chem. Eur. J. 10,
2555–2564.
35. Ellern, A., Bernstein, J., Becker, J. Y., Zamir, S., Shahal, L. & Cohen, S. (1994)
Chem. Mater. 6, 1378–1385.
36. Ashton, P. R., Balzain, V., Becher, J., Credi, A., Fyfe, M. C. T., Mattersteig,
G., Menzer, S., Nielsen, M. B., Raymo, F., M., Stoddart, J. F., et al. (1999) J. Am.
Chem. Soc. 121, 3951–3957.
37. Odell, B., Reddington, M. V., Slawin, A. M. Z., Spencer, N., Stoddart, J. F. &
Williams, D. J. (1988) Angew. Chem. Int. Ed. 27, 1547–1550.
38. Philp, D., Slawin, A. M. Z., Spencer, N., Stoddart, J. F. & Williams, D. J. (1991)
Chem. Commun. 22, 1584–1586.
Brough et al.
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