Polymer Translocation Research Articles

Investigating binding particles distribution effects on polymer translocation through nanopore

Chaperone driven polymer translocation is an important model for biopolymer's translocation in vivo. Binding proteins spatial distribution is a significant factor in calculating the translocation time of the polymer in this type of translocation. Here using a dynamical Monte Carlo simulation we compare the results of the usual uniform distribution with the exponential distribution of different rates for a stiff polymer. Our simulation results show that just by changing the chaperones spatial distribution the translocation time of the biopolymer will change by as large as an order. It can change the translocation regime of the polymer completely from a diffusive to a ballistic one. Although generally increasing the exponential rate and the background concentration will increase the translocation velocity, it is not always true and one should consider both the sequence and the background concentration. We show that the results depend on the sequence and changing the distribution rates for increasing the translocation velocity will change the whole Probability Density Function (PDF) of the polymer translocation time accordance to its sequence. The translocation time sequence dependency will change in the extreme cases e.g. in the high exponential rate. Investigating the binding protein size, λ, also shows the importance of the so called parking lot effect in distribution dependency of the translocation velocity. Although there is not any important dependency for λ=1, translocation time depends clearly on the chaperone spatial distribution for the case of λ≥2.

Chaperone-assisted translocation of flexible polymers in three dimensions.

Polymer translocation through a nanometer-scale pore assisted by chaperones binding to the polymer is a process encountered in vivo for proteins. Studying the relevant models by computer simulations is computationally demanding. Accordingly, previous studies are either for stiff polymers in three dimensions or flexible polymers in two dimensions. Here, we study chaperone-assisted translocation of flexible polymers in three dimensions using Langevin dynamics. We show that differences in binding mechanisms, more specifically, whether a chaperone can bind to a single site or multiple sites on the polymer, lead to substantial differences in translocation dynamics in three dimensions. We show that the single-binding mode leads to dynamics that is very much like that in the constant-force driven translocation and accordingly mainly determined by tension propagation on the cis side. We obtain β≈1.26 for the exponent for the scaling of the translocation time with polymer length. This fairly low value can be explained by the additional friction due to binding particles. The multiple-site binding leads to translocation the dynamics of which is mainly determined by the trans side. For this process we obtain β≈1.36. This value can be explained by our derivation of β=4/3 for constant-bias translocation, where translocated polymer segments form a globule on the trans side. Our results pave the way for understanding and utilizing chaperone-assisted translocation where variations in microscopic details lead to rich variations in the emerging dynamics.

Flow-induced translocation of star polymers through a nanopore

We study the flow-induced translocation of the star polymers through a nanopore using a hybrid simulation method that incorporates a lattice-Boltzmann approach for the fluid into a molecular dynamics model for the polymer. Our simulation demonstrates the existence of an optimal forward arm number of the star polymers captured by the nanopore, and illustrates its significance in determining the critical velocity flux of the star polymer translocation through the nanopore. Importantly, we find that the critical velocity flux of the star polymers is independent of the arm polymerization degree, but exhibits a linear dependence on the arm number. Based on previous scaling arguments and our simulation results, we conclude a linear dependence of the critical velocity flux on the arm number of the star polymers, which can successfully describe the dynamics of the star polymer translocation. Our simulation results rationalize the experimental results for the dependence of the critical velocity flux on the arm polymerization degree and the arm number of the star polymers, which provide new insights for the characterization and the purification of the star polymers.

Simulation of translocating pore of DNA in non-uniform force by coarse-grained model

Translocating pore of biomacromolecules is a common phenomenon in many biological processes, such as DNA transcription, cell infection of virus and transmembrane of proteins. The understanding of translocating pore of DNA is important for studying the DNA sequencing, gene therapy and virus infection. According to the coarse-grained model, we use molecular dynamics simulations to investigate the process of translocating pore of DNA under the actions of different non-uniform forces. In the present study, we consider five kinds of non-uniform forces, i.e., linearly increasing, linearly decreasing, V-type, inverted V-shaped, and periodic type. In the simulations of coarse-grained DNA, we find that the force on the pore opening palys a key role in the process of translocation of polymer. When the force is small, the probability of successful translocation of DNA is low accordingly. In the case of inverted V-shaped potential, the difference between the maximum and minimum force should be in a limited range to a probable translocation of DNA. Out of the range it might lead to clogged pores in the polymer chain. In the action of a non-uniform force, the translocating pore of DNA shows a series of complicated behaviors. For example, the end of a polymer can move faster than its head, resulting in the hole clogging and accumulation of polymers. A reversion can occasionally occur after a successful translocation of polymer. Therefore, non-uniform force leads to various scenarios of translocating pore of polymers.In summary, due to the complicated interactions between external forces and internal potential of polymer chains, particles can be clogged in the pore since the following particles overtake the leading ones in the chain. It is also found that the success of pore translation of DNA is significantly dependent on the acting force on the pore. Among all the cases of translating the pore successfully, the translation time in the case of non-uniform force is about half that in the case of uniform force. These results might provide an insight into the understanding of the complicated pore translating mechanism of DNA.

What controls open-pore and residual currents in the first sensing zone of alpha-hemolysin nanopore? Combined experimental and theoretical study.

The electrophoretic transport of single-stranded DNA through biological nanopores such as alpha-hemolysin (αHL) is a promising and cost-effective technology with the potential to revolutionize genomics. The rational design of pores with the controlled polymer translocation rates and high contrast between different nucleotides could improve significantly nanopore sequencing applications. Here, we apply a combination of theoretical and experimental methods in an attempt to elucidate several selective modifications in the pore which were proposed to be central for the effective discrimination between purines and pyrimidines. Our nanopore test set includes the wild type αHL and six mutants (E111N/M113X/K147N) in which the cross-section and chemical functionality of the first constriction zone of the pore are modified. Electrophysiological recordings were combined with all-atom Molecular Dynamics simulations (MD) and a recently developed Brownian Dynamics (BROMOC) protocol to investigate residual ion currents and pore-DNA interactions for two homo-polymers e.g. poly(dA)40 or poly(dC)40 blocking the pore. The calculated residual currents and contrast in the poly(dA)40/poly(dC)40 blocked pore are in qualitative agreement with the experimental recordings. We showed that a simple structural metric allows rationalization of key elements in the emergent contrast between purines and pyrimidines in the modified αHL mutants. The shape of the pore and its capacity for hydrogen bonding to a translocated polynucleotide are two essential parameters for contrast optimization. To further probe the impact of these two factors in the ssDNA sensing, we eliminated the effect of the primary constriction using serine substitutions (i.e. E111S/M113S/T145S/K147S) and increased the hydrophobic volume of the central residue in the secondary constriction (L135I). This pore modification sharply increased the contrast between Adenine (A) and Cytosine (C).

Polymer translocation through nanopore under external electric field: dissipative particle dynamics study

The DNA sequencing technology has achieved a leapfrog development in recent years. As a new generation of the DNA sequencing technology, nanopore sequencing has shown a broad application prospect and attracted vast research interests since it was proposed. In the present study, the dynamics of the electric-driven translocation of a homopolymer through a nanopore is investigated by the dissipative particle dynamics (DPD), in which the homopolymer is modeled as a worm-like chain (WLC). The DPD simulations show that the polymer chain undergoes conformation changes during the translocation process. The different structures of the polymer in the translocation process, i.e., single-file, double folded, and partially folded, and the induced current blockades are analyzed. It is found that the current blockades have different magnitudes due to the polymer molecules traversing the pore with different folding conformations. The nanoscale vortices caused by the concentration polarization layers (CPLs) in the vicinity of the sheet are also studied. The results indicate that the translocation of the polymer has the effect of eliminating the vortices in the polyelectrolyte solution. These findings are expected to provide the theoretical guide for improving the nanopore sequencing technique.

Translocation of a semiflexible polymer through a nanopore in the presence of attractive binding particles.

We study the translocation dynamics of a semiflexible polymer through a nanopore from the cis into the trans compartment containing attractive binding particles (BPs) using the Langevin dynamics simulation in two dimensions. The binding particles accelerate the threading process in two ways: (i) reducing the back-sliding of the translocated monomer, and (ii) providing the pulling force toward the translocation direction. We observe that for certain binding strength (ε_{c}) and concentration (ρ) of the BPs, the translocation is faster than the ideal ratcheting condition as elucidated by Simon, Peskin, and Oster [M. Simon, C. S. Peskin, and G. F. Oster, Proc. Natl. Acad. Sci. USA 89, 3770 (1992)PNASA60027-842410.1073/pnas.89.9.3770]. The asymmetry produced by the BPs at the trans-side leads to similarities of this process to that of a driven translocation with an applied force inside the pore manifested in various physical quantities. Furthermore, we provide an analytic expression for the force experienced by the translocating chain as well as for the scaled mean first passage time (MFPT), for which we observe that for various combinations of N,ε, and ρ the scaled MFPT (〈τ〉/N^{1.5}ρ^{0.8}) collapses onto the same master plot. Based on the analysis of our simulation data, we provide plausible arguments with regard to how the scaling theory of driven translocation can be generalized for such a directed diffusion process by replacing the externally applied force with an effective force.

Surface-adsorption-induced polymer translocation through a nanopore: Effects of the adsorption strength and the surface corrugation.

The surface corrugation plays an important role in single polymer diffusion on attractive surfaces. However, its effect on dynamics of surface adsorption-induced polymer translocation through a nanopore is not clear. Using three-dimensional Langevin dynamics simulations, we investigate the dynamics of a flexible polymer chain translocation through a nanopore induced by the selective adsorption of translocated segments onto the trans side of the membrane. The translocation probability Ptrans increases monotonically, while the mean translocation time τ has a minimum as a function of the adsorption strength ɛ, which are explained from the perspective of the effective driving force for the translocation. With the surface being smoother, τ as well as the scaling exponent α of τ with the chain length N decreases. Finally, we show that the distributions of the translocation time are non-Gaussian even for strong adsorption at a moderate surface corrugation. A nearly Gaussian distribution of the translocation time is observed only for the smoothest surface we studied.

Theory of polymer translocation through a flickering nanopore under an alternating driving force.

We develop a theory for polymer translocation driven by a time-dependent force through an oscillating nanopore. To this end, we extend the iso-flux tension propagation theory [Sarabadani et al., J. Chem. Phys. 141, 214907 (2014)] for such a setup. We assume that the external driving force in the pore has a component oscillating in time, and the flickering pore is similarly described by an oscillating term in the pore friction. In addition to numerically solving the model, we derive analytical approximations that are in good agreement with the numerical simulations. Our results show that by controlling either the force or pore oscillations, the translocation process can be either sped up or slowed down depending on the frequency of the oscillations and the characteristic time scale of the process. We also show that while in the low and high frequency limits, the translocation time τ follows the established scaling relation with respect to chain length N0, in the intermediate frequency regime small periodic, fluctuations can have drastic effects on the dynamical scaling. The results can be easily generalized for non-periodic oscillations and elucidate the role of time dependent forces and pore oscillations in driven polymer translocation.

Flow-Induced Ring Polymer Translocation through Nanopores

We investigate the flow-induced linear and ring polymer translocation through nanopores using a hybrid simulation method that combines a lattice-Boltzmann approach for the fluid with a molecular dynamics model for the polymer chain. Our results illustrate that the critical velocity flux for the linear and ring chain exhibits characteristic differences: for the linear chain, the critical velocity flux is independent of the chain length, which is in agreement with previous work; whereas, for the ring chain, the critical velocity flux decreases slowly with the increase of the chain length and gets closer to the critical velocity flux of the linear chain. In addition, we find that the ring chain shows much faster translocation compared to the linear chain with the same chain length. Moreover, the ring chain and the folded linear chain display the similar configurational deformation during the translocation. These results above indicate that with the increase of the chain length, the separation between the linear and ring polymers in the flow-induced translocation through nanopores becomes increasingly difficult.

Translocation of polymers into crowded media with dynamic attractive nanoparticles.

The translocation of polymers through a small pore into crowded media with dynamic attractive nanoparticles is simulated. Results show that the nanoparticles at the trans side can affect the translocation by influencing the free-energy landscape and the diffusion of polymers. Thus the translocation time τ is dependent on the polymer-nanoparticle attraction strength ɛ and the mobility of nanoparticles V. We observe a power-law relation of τ with V, but the exponent is dependent on ɛ and nanoparticle concentration. In addition, we find that the effect of attractive dynamic nanoparticles on the dynamics of polymers is dependent on the time scale. At a short time scale, subnormal diffusion is observed at strong attraction and the diffusion is slowed down by the dynamic nanoparticles. However, the diffusion of polymers is normal at a long time scale and the diffusion constant increases with the increase in V.

Effect of hydrodynamic interaction on flow-induced polymer translocation through a nanotube

We investigated the effect of hydrodynamic interaction(HI) on flow-induced polymer translocation through a nanotube by Brownian dynamics simulations. Whether there is HI in the simulation system is separately controlled by using different diffusion tensors. It is found that HI has no effect on critical velocity flux for long polymer chains due to the competition between more drag force and the hindrance of chain stretching from HI, however, HI broadens the transition interval. In addition, for flow-induced polymer translocation with HI, the critical velocity flux firstly slowly decreases with the increase of chain length and then becomes identical to that of it without HI, that is, the critical velocity flux is independent of chain length. At the same time, HI also accelerates the translocation process and makes the relative variation amplitude of single bead translocation time smaller. In fact, HI can enhance the intrachain cooperativity to make the whole chain obtain more drag force from fluid field and hinder chain stretching, both of which play an important role in translocation process.

Dynamic Scaling Theory of the Forced Translocation of a Semi-flexible Polymer Through a Nanopore

We present a theoretical description of the dynamics of a semi-flexible polymer being pulled through a nanopore by an external force acting at the pore. Our theory is based on the tensile blob picture of Pincus in which the front of the tensile force propagates through the backbone of the polymer, as suggested by Sakaue and recently applied to study a completely flexible polymer with self-avoidance, by Dubbledam et al. For a semi-flexible polymer with a persistence length P , its statistics is self-avoiding for a very long chain. As the local force increases, the blob size starts to decrease. At the blob size P/a^2 , where a is the size of a monomer, the statistics becomes that of an ideal chain. As the blob size further decreases to below the persistence length P, the statistics is that of a rigid rod. We argue that semi-flexible polymer in translocation should include the three regions: a self-avoiding region, an ideal chain region and a rigid rod region, under uneven tension propagation, instead of a uniform scaling picture as in the case of a completely flexible polymer. In various regimes under the effect of weak, intermediate and strong driving forces we derive equations from which we can calculate the translocation time of the polymer. The translocation exponent is given by \alpha=1+\mu, where \mu is an effective exponent for the end-to-end distance of the semi-flexible polymer, having a value between 1/2 and 3/5, depending on the total contour length of the polymer. Our results are of relevance for forced translocation of biological polymers such as DNA through a nanopore.

Polymer translocation into and out of an ellipsoidal cavity.

Monte Carlo simulations are used to study the translocation of a polymer into and out of an ellipsoidal cavity through a narrow pore. We measure the polymer free energy F as a function of a translocation coordinate, s, defined to be the number of bonds that have entered the cavity. To study polymer insertion, we consider the case of a driving force acting on monomers inside the pore, as well as monomer attraction to the cavity wall. We examine the changes to F(s) upon variation in the shape anisometry and volume of the cavity, the polymer length, and the strength of the interactions driving the insertion. For athermal systems, the free energy functions are analyzed using a scaling approach, where we treat the confined portion of the polymer to be in the semi-dilute regime. The free energy functions are used with the Fokker-Planck (FP) equation to calculate mean translocation times, as well as translocation time distributions. We find that both polymer ejection and insertion are faster for ellipsoidal cavities than for spherical cavities. The results are in qualitative agreement with those of a Langevin dynamics study in the case of ejection but not for insertion. The discrepancy is likely due to out-of-equilibrium conformational behaviour that is not accounted for in the FP approach.

Ionic current inversion in pressure-driven polymer translocation through nanopores.

We predict streaming current inversion with multivalent counterions in hydrodynamically driven polymer translocation events from a correlation-corrected charge transport theory including charge fluctuations around mean-field electrostatics. In the presence of multivalent counterions, electrostatic many-body effects result in the reversal of the DNA charge. The attraction of anions to the charge-inverted DNA molecule reverses the sign of the ionic current through the pore. Our theory allows for a comprehensive understanding of the complex features of the resulting streaming currents. The underlying mechanism is an efficient way to detect DNA charge reversal in pressure-driven translocation experiments with multivalent cations.

Dynamics of polymer translocation through kinked nanopores.

Polymer translocation through nanopore has potential technological applications for DNA sequencing, where one challenge problem is to slow down translocation speed. Inspired by experimental findings that kinked nanopores exhibit a large reduction in translocation velocity compared with their straight counterparts, we investigate the dynamics of polymer translocation through kinked nanopores in two dimensions under an applied external field. With increasing the tortuosity of an array of nanopores, our analytical results show that the translocation probability decreases. Langevin dynamics simulation results support this prediction and further indicate that with increasing the tortuosity, translocation time shows a slow increase followed by a rapid increase after a critical tortuosity. This behavior demonstrates that kinked nanopores can effectively reduce translocation speed. These results are interpreted by the roles of the tortuosity for decreasing the effective nanopore diameter, increasing effective nanopore length, and greatly increasing the DNA-pore friction.

A coarse‐grained MARTINI‐like force field for DNA unzipping in nanopores

In nanopore force spectroscopy (NFS) a charged polymer is threaded through a channel of molecular dimensions. When an electric field is applied across the insulating membrane, the ionic current through the nanopore reports on polymer translocation, unzipping, dissociation, and so forth. We present a new model that can be applied in molecular dynamics simulations of NFS. Although simplified, it does reproduce experimental trends and all-atom simulations. The scaled conductivities in bulk solution are consistent with experimental results for NaCl for a wide range of electrolyte concentrations and temperatures. The dependence of the ionic current through a nanopore on the applied voltage is symmetric and, in the voltage range used in experiments (up to 2 V), linear and in good agreement with experimental data. The thermal stability and geometry of DNA is well represented. The model was applied to simulations of DNA hairpin unzipping in nanopores. The results are in good agreement with all-atom simulations: the scaled translocation times and unzipping sequence are similar.

Active polymer translocation in the three-dimensional domain.

In this work we study the translocation process of a polymer through a nanochannel where a time dependent force is acting. Two conceptually different types of driving are used: a deterministic sinusoidal one and a random telegraph noise force. The mean translocation time presents interesting resonant minima as a function of the frequency of the external driving. For the computed sizes, the translocation time scales with the polymer length according to a power law with the same exponent for almost all the frequencies of the two driving forces. The dependence of the translocation time with the polymer rigidity, which accounts for the persistence length of the molecule, shows a different low frequency dependence for the two drivings.

A Molecular Description of Cellulose Biosynthesis

Cellulose is the most abundant biological macromolecule and is an extracellular linear polymer of glucose molecules. It is an essential component of plant cell walls but is also found in algae and bacteria. In bacteria, cellulose production frequently correlates with the formation of biofilms, a sessile, multi-cellular growth form. Cellulose synthesis and transport across the inner bacterial membrane is mediated by a complex of the multi-spanning catalytic BcsA subunit and the membrane-anchored periplasmic BcsB protein. We determined several crystal structures of the BcsA-B complex at intermediate states during cellulose synthesis and membrane translocation. The structures demonstrate how BcsA forms a cellulose-conducting channel and delineate conformational changes of the synthase underlying substrate binding, glycosyl transfer and polymer translocation. Furthermore, combining biochemical and structural data, we reveal the mechanism by which cyclic-di-GMP, a potent stimulator of bacterial biofilms and allosteric activator of bacterial cellulose synthase, regulates BcsA's activity.

Iso-flux tension propagation theory of driven polymer translocation: the role of initial configurations.

We investigate the dynamics of pore-driven polymer translocation by theoretical analysis and molecular dynamics (MD) simulations. Using the tension propagation theory within the constant flux approximation we derive an explicit equation of motion for the tension front. From this we derive a scaling relation for the average translocation time τ, which captures the asymptotic result τ∝N0(1+ν), where N0 is the chain length and ν is the Flory exponent. In addition, we derive the leading correction-to-scaling term to τ and show that all terms of order N0(2ν) exactly cancel out, leaving only a finite-chain length correction term due to the effective pore friction, which is linearly proportional to N0. We use the model to numerically include fluctuations in the initial configuration of the polymer chain in addition to thermal noise. We show that when the cis side fluctuations are properly accounted for, the model not only reproduces previously known results but also considerably improves the estimates of the monomer waiting time distribution and the time evolution of the translocation coordinate s(t), showing excellent agreement with MD simulations.