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Model of Inversion of DNA Charge by a Positive Polymer: Fractionalization of the Polymer Charge
T. T. Nguyen and B. I. Shklovskii
Theoretical Physics Institute, University of Minnesota, 116 Church St. Southeast, Minneapolis, Minnesota 55455 (Received 11 September 2001; published 17 June 2002) We model one strand of DNA by a one-dimensional lattice (ODL) of negative charges and consider the problem of inversion of its charge by a positive polyelectrolyte (PE). In the neutral state of the ODL-PE complex, each of the ODL charges is locally compensated by a PE charge. When an additional PE molecule is adsorbed by ODL, its charge gets fractionalized into monomer charges of defects (tails and arches) on the background of the perfectly neutralized ODL. Defects spread all over the ODL, eliminating the self-energy of PE. For DNA this fractionalization mechanism leads to a substantial inversion of charge, a phenomenon which is widely used for gene delivery.
DOI: 10.1103/PhysRevLett.89.018101 PACS numbers: 87.14.Gg, 87.15.Nn, 87.16.Dg

The inversion of the negative charge of a DNA double helix by its complexation with a positive polyelectrolyte (PE) is used for gene delivery. The positive charge of a DNA-PE complex facilitates DNA contact with a typically negative cell membrane making penetration into the cell hundreds times more likely [1]. Charge inversion of DNA-PE complexes was conrmed recently by electrophoresis [2]. At a given concentration of long DNA helices, when the concentration of shorter PE molecules increases, at some critical concentration the electrophoretic mobility of a DNA-PE complex changes sign from negative to positive. The challenging and counterintuitive phenomenon of charge inversion of a macroion by oppositely charged PE and other multivalent ions has attracted signicant attention [313]. Intuitively, one can think that, when a PE completely neutralizes a large macroion such as the DNA double helix, new molecules of a PE do not attach to the macroion. Indeed, the Poisson-Boltzmann approximation for the description of screening of a macroion by any counterions including PE does not lead to charge inversion. Charge inversion can be explained if one takes into account that the surface potential of an already-neutralized macroion is locally affected by a new approaching PE molecule or, in other words, it can be explained if one takes into account correlations between PE molecules [9,13]. For quantitative consideration, the charges of a macroion are always assumed to be uniformly smeared. This approach ignores the interference between the structure of the macroion surface and that of a PE and clearly is not fully satisfactory. More importantly, it is not clear whether or not charge inversion is an artifact of the assumption of uniformly smeared charge. In this paper, we consider the effects of discreteness of 2e charges of the macroion. We show that in this case charge inversion not only exists but can be explained as originating from a fractionalization of charge of PE molecules. This explanation turns out to be even simpler and more visual than for the model of smeared charges of the macroion. 018101-1 0031-9007 02 89(1) 018101(4)$20.00

To discuss the mechanism of fractionalization, we use a model of DNA as an example. In DNA, negative elementary charges of phosphate groups are situated along the two spirals at the exterior of both strands. When unfolded, each spiral is a one-dimensional lattice of such charges, with the 6.7 . To study the fractionalization lattice constant a mechanism of charge inversion, we replace a DNA strand by a model of a rigid, one-dimensional lattice (ODL) of negative charges with lattice constant a. We also consider a model of a PE as a freely jointed chain of Z small 1e monomers. To maximize the role of discreteness of ODL charges we begin with the assumption that the PE bond length b is exactly equal to the lattice constant a of the ODL. We call such PE matching. We also assume that the distance of closest approach, d, between a PE charge and a ODL charge is smaller than a. Then PE molecules can attach to a ODL in such a way that every charge of the ODL is locally compensated by a PE charge and, therefore, the ODL is completely neutralized. The case of Z 3 is shown in Fig. 1a. The neutralization by a matching PE is so perfect that it is difcult to imagine how another PE

FIG. 1. The origin of charge fractionalization. (a) ODL of negative charges (empty circles) is completely neutralized by positive PE molecules with Z 3 (their charges are shown by solid circles). A new PE molecule is approaching. (b) The new PE molecule is digested, its charge is split into 1e charges of two tails and an arch.

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molecule can be attached to the ODL. Thus, it seems to be impossible to overcharge the ODL. In this paper, we, however, show that, even in this worst scenario for charge inversion, there is a mechanism which brings an additional PE to the neutralized ODL and leads to its charge inversion. We call this mechanism fractionalization, and Fig. 1 shows how this mechanism works 3. When a new PE comes to the alfor the case of Z ready-neutralized ODL, it creates a place for itself or, in other words, the oppositely charged image on the ODL in the following way. One PE monomer from Z alreadyadsorbed PE molecules detaches from the ODL surface. This leads to the formation of Z positive defects (tails and arches) and Z negative vacancies on the ODL. All Z vacancies can then join together and form a large vacancy of length Z by shifting adsorbed PE molecules along ODL. A new PE molecule is accommodated in this vacancy. As a result of the consumption of this molecule, Z defects, each with charge 1e, appear on the top of the completely neutralized ODL (see Fig. 1b). This effectively looks like cutting the new PE molecule into Z individual monomers and spreading them out along the ODL. In other words, charge inversion of ODL happens by fractionalization of the PE molecule charge. Of course, none of the chemical bonds are really cut, and this phenomenon is solely due to the correlated distribution of PE molecules, which avoid each other at the ODL. In this sense, the fractionalization we are talking about is similar to what happens in a fractional quantum Hall effect [14] or in the polyacytilene [15], where many-electron correlations result in the effective fractionalization of the electron charge, while no electrons are actually split. It is also similar to the spin-charge seperation into holon and spinon in a one-dimensional antiferromagnet with a single hole. We believe that we have found a valuable addition to these beautiful and rare effects. Fractionalization is driven by the elimination of the selfenergy of free PE molecules. By self-energy, we mean the energy of repulsive interactions of Z positive charges of the PE molecule in the extended conformation which it has in the bulk solution. In the fractionalized state, charges of defects are very far from each other and practically do not interact, so that the positive PE self-energy is eliminated. How does fractionalization work when distance between charges of PE, b, is not equal to the distance between charges of the ODL, a? Consider, for example, commensurate PE with b a 2, which has twice as large linear density of charge than a ODL. In this case, due to their strong Coulomb repulsion, PE molecules form an analog of Wigner crystal, where PE molecules alternate with vacant places (see Fig. 2a). In this case, even if the PE is not exible a new PE molecule creates Z distant grain boundaries (domain walls), where one vacancy is missing (see Fig. 2b). The charge of each grain boundary is 1e, so that charge of the new PE molecule is fractionalized and most of the self-energy of PE is eliminated in the way similar to what happens in the case of matching PE. 018101-2

FIG. 2. The origin of charge fractionalization for a PE with linear charge density twice as large as that for the ODL. (a) A Wigner-crystal-like ground state of a periodic chain of negative charges (empty circles) neutralized by PE molecules with Z 3 and b a 2 (their monomers are shown by solid circles). (b) The new PE molecule is digested by the ODL. Its charge is split into 1e charges of Z grain boundaries.

Fractionalization continues to work when the linear charge density of a multivalent counterion (Z ion) is even larger. We can imagine such a limit, when dealing with a metallic multivalent ion (for example, La13 ), which touches only one negative charge of ODL. In this case, the ground state of the neutralized ODL resembles a Wigner crystal even closer (Fig. 3a). Fractionalization of the charge of a new PE molecule into Z monovalent charges of grain boundaries (see Fig. 3b) partially eliminates its self-energy and drives charge inversion. Figure 3 is actually a very crude picture [14] of the fractional Hall effect at lling factor 1 3. In the latter case, the empty circles represent discrete Landau states and an electron charge 2e is split into three charges 2e 3 (the positive charge is provided by the background of donors, not by Landau states). Figure 3 also resembles what happens in the case of the adsorption of Z ions on the line or surface with uniform distribution of background charge [9,10]. In that case, the charge of the new Z ion is smeared along the background due to small elastic deformations of Wigner-crystal-like strongly correlated liquid. In other words, the Z ion is fractionalized into innitesimally small portions. One can visualize the transition to the case of uniform surface charge,

FIG. 3. The origin of charge fractionalization for multivalent counterions. (a) A Wigner-crystal-like ground state of a ODL of negative charges (empty circles) neutralized by multivalent counterions with Z 3 (larger solid circles). (b) The new Z ion is digested by the ODL. Its charge is split into 1e charges of Z grain boundaries.

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imagining that both elementary charges of our lattice and lattice constant a vanish, while the charge density of ODL and the charge of the Z ion are kept constant. Let us return to the adsorption of PE with a nite linear charge density on ODL and discuss the situations where b , a but b and a are not commensurable. Even in this case the ground state of a neutralized ODL is a crystal. If an additional PE molecule is adsorbed it is still fractionalized to Z grain boundaries with charge 1e. The only difference from the commensurate case shown in Fig. 2 is that grain boundary can be more complicated and include several PE molecules. It is well known that fractionalization of charge of Z ions into free grain boundaries shown in Fig. 3 cannot be generalized to a two-dimensional case because of the costly creation of linear defects (strings) which connect two pointlike defects. However, for a exible matching PE, fractionalization into free tails and arches is not limited by one dimension. It is easy to see from Fig. 4 that the same mechanism applies equally well to a two-dimensional square lattice of discrete negative charges with the lattice constant a equal to the PE bond length b. All previous arguments about the role of tails and arches for ODL can be carried over to this two-dimensional case. There are no strings between defects in this case. This is a remarkable consequence of the involvement of the third dimension. We do not know any other classical example of charge fractionalization in a two-dimensional system, which has no strong uniaxial anisotropy. Two-dimensional fractionalization and the resulting charge inversion are of course also driven by self-energy elimination. An important role of elimination of the self-energy for the adsorption of a exible PE on an opposite uniformly charged surface can be traced in Refs. [4,6,12]. Let us now demonstrate how the physical idea of the fractionalization can be used to estimate the net inverted charge of a DNA. Because, the DNA molecule is a double helix, we model it primitively as a rigid cylinder with

radius R which has two ODLs of negative charges spiraling on its surface. Let us denote the linear charge density of the inverted (positive) net charge of the cylinder by h . It includes both linear density of the bare charge, h0 , of the cylinder and the charge of adsorbed PE molecules. The chemical potential of the PE absorbed at the spiral is ms ZkB T ln h h0 1 Zec 0 . (1)

The rst term on the right-hand side of Eq. (1) is the chemical potential of the one-dimensional gas of defects. We used the expression for the chemical potential of an ideal gas because the Coulomb interaction energy between defects at a distance of a few a is much smaller than kB T (a lB , where lB e2 DkB T 7 is the Bjerrum length, and D is the dielectric constant of water). The second term on the right-hand side of Eq. (1) is the interaction energy of the new PE molecule with the inverted charge of the cylinder. In this term, c 0 is the averaged surface potential of the cylinder. We assume in this paper that the cylinder net charge is screened by a monovalent salt at the screening length rs , which is much larger than a. Then the potential c 0 is that of a uniformly charged cylinder with radius R and linear charge density h : c 0 2h rs 1 R ln . D R (2)

To nd h in the equilibrium state, one has to equate the chemical potential of adsorbed PE molecules with that of a free PE in the solution. The latter one can be calculated as follows. Because of the repulsive Coulomb interaction between monomers, a free PE in the solution has an extended shape to minimize its energy. Therefore, the chemical potential of a free PE in solution can be written as the self-energy of a rigid rod with the length Za and the linear charge density e a, m0 Ze2 Da ln L a , (3)

+ +

where L min rs , Za . We have assumed that the concentration of PE in the solution is large enough so that its translational entropy can be neglected. In this sense, we are calculating the maximum possible charge inversion of the cylinder. If the PE molecule is long Z 1 this limit is already reached at a concentration of PE which is exponentially small exp 2Z . Equating the chemical potentials of Eqs. (3) and (1), one has c 0 e Da ln L a 1 kB T e ln h0 h . (4)

FIG. 4. Fractionalization of an additional PE molecule with Z 3 into three positive defects at a two-dimensional lattice of negative charges (white spheres) neutralized by alreadyadsorbed PE. Positive charges of PE molecules are shown by black spheres.

One can interpret the right-hand side as a correlation voltage that (over)charges the cylinder to the potential c 0. To the rst approximation, one can neglect the entropic term on the right-hand side of Eq. (4) and easily obtain a solution for the net charge density h e ln L a . 2a ln rs 1 R R (5) 018101-3

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Now one can check that this solution is consistent with the assumption that the entropic term can be neglected by substituting it back into Eq. (4). Of course, h is positive, indicating that the cylinder charge is inverted. Knowing 0.6e 3.9e a, we obtain h and using jh0 j ln L a h 0.13 . (6) h0 ln rs 1 R R For an example of the DNA double helix, R 10 and a 6.7 , so that at rs $ 10 the ratio of logarithms can be only slightly larger than unity. Thus, the charge inversion ratio created by fractionalization is limited by 20%. Up to such a point we indeed can neglect Coulomb interactions between defects in the chemical potential of the gas of defects [the rst term on the right-hand side of Eq. (1)]. To conclude, the main result of this paper is that discreteness of surface charges of a macroion, for example, a double helix DNA, does not prevent its overcharging by oppositely charged PE and other Z ions. Even in the worst scenario of matching PE and macroion geometries, when in the neutral state all charges of macroions are perfectly neutralized by PE, charge inversion happens due to fractionalization of PE charge into 1e charges of defects. This is an extremely transparent mechanism, as illustrated by Figs. 1 and 4. In the nonmatching cases the mechanism of charge inversion for discrete surface charges looks more similar to the one previously discussed in a model of a uniformly charged macroion surface [9,13], but is still accompanied by fractionalization. We emphasize that charge inversion always happens due to the fact that a new PE molecule rearranges alreadyadsorbed PE in such a way that its image strongly attracts this new PE molecule and the self-energy of the new PE is (partially) eliminated. For an example of the fractionalization mechanism studied in this paper, in Fig. 1, a three-site vacancy for a new PE molecule is created due to the creation of three defects with charge e each. When the new molecule binds to its image (occupies the vacancy), its self-energy is eliminated. This physics of self-energy elimination (or attraction to the image) cannot be described by the Poisson-Boltzmann theory because this theory uses the mean-eld potential, which does not depend on the position of a new PE molecule. We are grateful to A. V. Kabanov, whose question initiated this work, and A. Yu. Grosberg, V. A. Kabanov, and P. Pincus for useful discussions of our results. This work was supported by an NSF grant No. DMR-9985785.

Note added. If a, R rs Za the ratio of logarithms in Eq. (5) equals unity and the linear density of net charge of DNA h is equal to half of the linear charge density of PE, hPE e a, i.e., h hPE 2 . (7)

This result follows just from the fact that practically all the self-energy of PE is eliminated. Therefore, it is not sensitive to the microscopic structure of DNA-PE complex. This is conrmed by the fact that parameters of DNA do not show up in Eq. (7). The nonmatching case shown in Fig. 2 is another example of the universality of Eq. (7). This is because most of the self-energy of PE is eliminated in this case as well. In a recent paper [16] the universality of Eq. (7) was conrmed for the same range a, R rs Za. The authors studied the model of DNA as a uniformly charge cylinder, on which PE is free to form a perfect coil with the optimal period, again eliminating most of its self-energy.

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