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Biomedical subjects

S A Allison

Publications and source records attributed to S A Allison.

At least 19 recordsLinked to original sources
PubMed2001

Boundary element modeling of biomolecular transport.

The boundary element (BE) methodology has emerged as a powerful tool in modeling a broad range of different transport phenomena of biomolecules in dilute solution. These include: sedimentation, diffusion (translational and rotational), intrinsic viscosity, and free solution electrophoresis. Modeling is carried out in the framework of the continuum primitive model where the biomolecule is modeled as an arbitrary array of solid platelets that contains fixed charges within. The surrounding fluid is modeled as a electrodynamic/hydrodynamic continuum which obeys the Poisson and low Reynolds number Navier-Stokes equations. Ion relaxation (the distortion of the ion atmosphere from equilibrium) can also be accounted for by solving the coupled ion transport equation (for each mobile ion species present), Poisson, and Navier-Stokes equations in tandem. Several examples are presented in this work. It is first applied to a detailed model of 20 bp DNA and it is concluded that it is not necessary to include a layer of bound water to reconcile experimental and model translational diffusion constants. With regards to diffusion, the BE approach is also applied to a 375-bp supercoiled DNA model (without ion relaxation), and also 20-60-bp DNA fragments with ion relaxation included in order to assess the magnitude of the electrolyte friction effect under a number of different salt/buffer conditions. Attention is then turned to modeling the electrophoretic mobility of three different cases. First of all, we consider a sphere with a central charge large enough in magnitude to insure that ion relaxation is significant. Excellent agreement with independent theory is obtained. Finally, it is applied to modeling short DNA fragments in KCl and Tris acetate salts. Quantitative agreement is achieved when the salt is KCl, but the calculated (absolute) mobility in Tris acetate is substantially higher than the experimental value. The interpretation of this is that there is an association between Tris(+) and DNA (perhaps hydrogen bonding) not accounted for in our modeling that is responsible for this discrepancy.

Biological Transport↗
PubMed2000

A commentary on the screened-Oseen, counterion-condensation formalism of polyion electrophoresis.

The use of linear theory, in particular, counterion condensation (CC) theory, in describing electrophoresis of polyelectrolyte chains, is criticized on several grounds. First, there are problems with CC theory in describing the equilibrium distribution of ions around polyelectrolytes. Second, CC theory is used to treat ion relaxation in a linear theory with respect to the polyion charge despite the fact that ion relaxation arises as a consequence of nonlinear charge effects. This nonlinearity has been well established by several investigators over the last 70 years for spherical, cylindrical, and arbitrarily shaped model polyions. Third, current use of CC theory ignores the electrophoretic hindrance as well as the ion relaxation for condensed counterions and only includes such interactions for uncondensed counterions. Because most of the condensed counterions lie outside the shear surface of the polyion (in the example of DNA), the assumption of ion condensation is artificial and unphysical. Fourth, the singular solution, based on a screened Oseen tensor, currently used in the above mentioned theories is simply wrong and fails to account for the incompressibility of the solvent. The actual singular solution, which has long been available, is discussed. In conclusion, it is pointed out that numerical alternatives based on classic electrophoresis theory (J.T.G. Overbeek, Kolloid-Beih, 1943, 54:287-364) are now available.

Biopolymers↗
PubMed1999

Visualizing ion relaxation in the transport of short DNA fragments.

Ion relaxation plays an important role in a wide range of phenomena involving the transport of charged biomolecules. Ion relaxation is responsible for reducing sedimentation and diffusion constants, reducing electrophoretic mobilities, increasing intrinsic viscosities, and, for biomolecules that lack a permanent electric dipole moment, provides a mechanism for orienting them in an external electric field. Recently, a numerical boundary element method was developed to solve the coupled Navier-Stokes, Poisson, and ion transport equations for a polyion modeled as a rigid body of arbitrary size, shape, and charge distribution. This method has subsequently been used to compute the electrophoretic mobilities and intrinsic viscosities of a number of model proteins and DNA fragments. The primary purpose of the present work is to examine the effect of ion relaxation on the ion density and fluid velocity fields around short DNA fragments (20 and 40 bp). Contour density as well as vector field diagrams of the various scalar and vector fields are presented and discussed at monovalent salt concentrations of 0.03 and 0.11 M. In addition, the net charge current fluxes in the vicinity of the DNA fragments at low and high salt concentrations are briefly examined and discussed.

Biophysical Phenomena↗
PubMed1997

Modeling the electrophoresis of lysozyme. II. Inclusion of ion relaxation.

In this work, boundary element methods are used to model the electrophoretic mobility of lysozyme over the pH range 2-6. The model treats the protein as a rigid body of arbitrary shape and charge distribution derived from the crystal structure. Extending earlier studies, the present work treats the equilibrium electrostatic potential at the level of the full Poisson-Boltzmann (PB) equation and accounts for ion relaxation. This is achieved by solving simultaneously the Poisson, ion transport, and Navier-Stokes equations by an iterative boundary element procedure. Treating the equilibrium electrostatics at the level of the full rather than the linear PB equation, but leaving relaxation out, does improve agreement between experimental and simulated mobilities, including ion relaxation improves it even more. The effects of nonlinear electrostatics and ion relaxation are greatest at low pH, where the net charge on lysozyme is greatest. In the absence of relaxation, a linear dependence of mobility and average polyion surface potential, (lambda zero)s, is observed, and the mobility is well described by the equation [formula: see text] where epsilon 0 is the dielectric constant of the solvent, and eta is the solvent viscosity. This breaks down, however, when ion relaxation is included and the mobility is less than predicted by the above equation. Whether or not ion relaxation is included, the mobility is found to be fairly insensitive to the charge distribution within the lysozyme model or the internal dielectric constant.

Electrophoresis↗
PubMed1995

Monte Carlo simulations of supercoiling free energies for unknotted and trefoil knotted DNAs.

A new Monte Carlo (MC) algorithm is proposed for simulating inextensible circular chains with finite twisting and bending rigidity. This new algorithm samples the relevant Riemann volume elements in a uniform manner, when the constraining potential vanishes. Simulations are performed for filaments comprising 170 subunits, each containing approximately 28 bp, which corresponds to a DNA length of 4770 bp. The bending rigidity is chosen to yield a persistence length, P = 500 A, and the intersubunit potential is taken to be a hard-cylinder potential with diameter d = 50 A. This value of d yields the same second virial coefficient as the electrostatic potential obtained by numerical solution of the Poisson-Boltzmann equation for 150 mM salt. Simulations are performed for unknotted circles and also for trefoil knotted circles using two different values of the torsional rigidity, C = (2.0 and 3.0) x 10(-19) dyne cm2. In the case of unknotted circles, the simulated supercoiling free energy varies practically quadratically with linking difference delta l. The simulated twist energy parameter ET, its slope dET/dT, and the mean reduced writhe /delta l for C = 3 x 10(-19) dyne cm2 all agree well with recent simulations for unknotted circles using the polygon-folding algorithm with identical P, d, and C. The simulated ET vs. delta l data for C = 2.0 x 10(-19) dyne cm2 agree rather well with recent experimental data for p30 delta DNA (4752 bp), for which the torsional rigidity, C = 2.07 x 10(-19) dyne cm2, was independently measured. The experimental data for p30 delta are enormously more likely to have arisen from C = 2.0 x 10(-19) than from C = 3.0 x 10(-19) dyne cm2. Serious problems with the reported experimental assessments of ET for pBR322 and their comparison with simulated data are noted. In the case of a trefoil knotted DNA, the simulated value, (ET)tre, exceeds that of the unknotted DNA, (ET)unk, by approximately equal to 1.40-fold at magnitude of delta l = 1.0, but declines to a plateau about 1.09-fold larger than (ET)unk when magnitude of delta l > or = 15. Although the predicted ratio, (ET)tre/(ET)unk approximately equal to 1.40, agrees fairly well with recent experimental measurements on a 5600-bp DNA, the individual measured ET values, like some of those reported for pBR322, are so large that they cannot be simulated using P = 500 A, d = 50 A, and any previous experimental estimate of C.

Algorithms↗
PubMed1995

Modeling the electrophoresis of rigid polyions: application to lysozyme.

An algorithm is developed to determine the electrophoretic mobility of a rigid polyion modeled as a low dielectric volume element of arbitrary shape containing an arbitrary charge distribution. The solvent is modeled as a high dielectric continuum with salt distributed according to the linearized Poisson Boltzmann equation. Account is also taken of a Stern layer that separates the molecular surface and the surface of hydrodynamic shear, or Stern surface. Relaxation of the ion atmosphere because of the presence of the external field is ignored. The electrostatic and hydrodynamic problems are both solved by boundary element methods. The procedure is first applied to spherical polyions containing monopolar, dipolar, and quadrupolar charge distributions, and calculated mobilities are found to be in excellent agreement with the theory of Yoon and Kim. It is then applied to lysozyme by using models that account for the detailed shape and charge distribution of the enzyme. For reasonable choices of the molecular and Stern surfaces, calculated and experimental mobilities are found to be in fair agreement with each other. However, if a pH independent Stern layer (or, equivalently, translational diffusion constant, Dt) is assumed, the calculated mobilities exhibit a stronger pH dependence than is observed experimentally. A small increase in Dt with increasing pH could correct this discrepancy.

Algorithms↗
PubMed1990

Point charge distributions and electrostatic steering in enzyme/substrate encounter: Brownian dynamics of modified copper/zinc superoxide dismutases.

The electrostatic steering mechanism of bovine erythrocyte Cu/Zn superoxide dismutase (SOD) was investigated through the use of Brownian dynamics. Simulations of enzyme/substrate encounter were carried out on 14 different SOD models defined by simple changes in the enzyme's point charge distribution. The magnitude and ionic strength dependence of reaction rates, rates for collision anywhere on the enzyme surface, and collision efficiency factors were analyzed to elucidate both the general and specific roles for point charges associated with amino acid residues. Collision rates for the general enzyme surface appear to be solely determined by the net charge on the enzyme. At physiological ionic strength this effect is negligible, with only 6% variation in collision rates observed as the net charge ranges from +2e to -10e. With the exception of a few charged residues in the active-site channel of SOD, point charge modifications had modest effects on reaction rates. For a large region within and surrounding the channel, reaction rates increased or decreased by only 10-15% with the addition or subtraction of a protonic unit of charge, respectively. This effect simply disappeared with increasing distance from the active site. More dramatic effects were seen at only three residues: arginine-141, glutamate-131, and lysine-134. Implications for rate enhancement through site-directed mutagenesis are discussed.

Amino Acids↗
PubMed1988

Amplification of bleomycin-mediated degradation of DNA by polyamines.

The degradation of calf thymus DNA by a ferrous complex of bleomycin A2 or Blenoxane (a mixture consisting mainly of BLM-A2 and BLM-B2) is enhanced by polyamines higher than ethylenediamine. The QSAR analysis gave excellent correlation between the experimental amplification results and calculated valence molecular connectivity indices of the third order and path type for the protonated polyamines. A new amplifier of bleomycin activity has been synthesized and its interaction with DNA has been studied. This compound contains a DNA-intercalating moiety and a polyamine portion, two independent amplification systems in the same molecule. The role of the C-terminus of bleomycin as the intramolecular amplifier for the degradation of DNA is discussed.

Animals↗
PubMed1986

Simulation of biomolecular diffusion and complex formation.

Diffusion is a phenomenon of very widespread importance in molecular biophysics. Diffusion can determine the rates and character of the assembly of multisubunit structures, the binding of ligands to receptors, and the internal motions of molecules and assemblies that involve solvent surface displacements. Current computer simulation techniques provide much more detailed descriptions of diffusional processes than have been available in the past. Models can be constructed to include such realistic features as structural subunits at the submolecular level (domains, monomers, or atoms); detailed electrostatic charge distributions and corresponding solvent-screened inter- and intramolecular interactions; and hydrodynamic interactions. The trajectories can be analyzed either to provide direct information on biomolecular function (e.g., the bimolecular rate constant for formation of an electron-transfer complex between two proteins), or to provide or test models for the interpretation of experimental data (e.g., the time dependence of fluorescence depolarization for segments of DNA). Here, we first review the theory of diffusional simulations, with special emphasis on new techniques such as those for obtaining transport properties of flexible assemblies and rate constants of diffusion-controlled reactions. Then we survey a variety of recent applications, including studies of large-scale motion in DNA segments and substrate "steering" in enzyme-substrate binding. We conclude with a discussion of current work (e.g., formation of protein complexes) and possible areas for future work.

Biopolymers↗