Biomedical subjects

Marcel Aguilella-Arzo

Publications and source records attributed to Marcel Aguilella-Arzo.

4 recordsLinked to original sources

PubMed2007

Electrostatic properties and macroscopic electrodiffusion in OmpF porin and mutants.

The bacterial porin OmpF found in the outer membrane of E. coli is a wide channel, characterized by its poor selectivity and almost no ion specificity. It has an asymmetric structure, with relatively large entrances and a narrow constriction. By applying continuum electrostatic methods we determine the ionization states of titratable amino acid residues in the protein and calculate self-consistently the electric potential 3-D distribution within the channel. The average electrostatic properties are then represented by an effective fixed charge distribution along the pore which is the input for a macroscopic electrodiffusion model. The theoretical predictions agree with measurements performed under different salt gradients and pH. The sensitivity of reversal potential and conductance to the direction of the salt gradient and the solution pH is captured by the model. The theory is also able to explain the influence of the lipid membrane charge. The same methodology is satisfactorily applied to some OmpF mutants involving slight structural changes but a large number of net charges. The correlation found between atomic structure and ionic selectivity shows that the transport characteristics of wide channels like OmpF and its mutants are mainly regulated by the collective action of a large number of residues, rather than by the specific interactions of residues at particular locations.

Amino Acid Substitution↗

PubMed2006

Blocking of an ion channel by a highly charged drug: modeling the effects of applied voltage, electrolyte concentration, and drug concentration.

We present a simple physical model to estimate the blocked pore probability of an ion channel that can be blocked by a highly charged drug in solution. The model is inspired by recent experimental work on the blocking of the PA(63) channel, involved in the anthrax toxin infection, by a highly charged drug [Karginov PNAS 102, 15075 (2005)]. The drug binding to the pore is highly specific but the strong dependence of blocking on the applied voltage and electrolyte concentration suggests that long range electrostatic interactions are important. Since basic electrostatic concepts rather than detailed molecular models are considered, the microscopic details of the channel blocking are ignored, although the model captures most of the qualitative characteristics of the problem.

Computer Simulation↗

PubMed2005

Computing numerically the access resistance of a pore.

The access resistance (AR) of a channel is an important component of the conductance of ion channels, particularly in wide and short channels, where it accounts for a substantial fraction of the total resistance to the movement of ions. The AR is usually calculated by using a classical and simple expression derived by Hall from electrostatics (J.E. Hall 1975 J. Gen. Phys. 66:531-532), though other expressions, both analytical and numerical, have been proposed. Here we report some numerical results for the AR of a channel obtained by solving the Poisson-Nernst-Planck equations at the entrance of a circular pore. Agreement is found between numerical calculations and analytical results from Hall's equation for uncharged pores in neutral membranes. However, for channels embedded in charged membranes, Hall's expression overestimates the AR, which is much lower and can even be neglected in some cases. The weak dependence of AR on the pore radius for charged membranes at low salt concentration can be exploited to separate the channel and the access contributions to the measured conductance.

Biological Transport↗

PubMed2004

Salting out the ionic selectivity of a wide channel: the asymmetry of OmpF.

Although the crystallographic structure of the bacterial porin OmpF has been known for a decade, the physical mechanisms of its ionic selectivity are still under investigation. We address this issue in a series of experiments with varied pH, salt concentrations, inverted salt gradient, and charged and uncharged lipids. Measuring reversal potential, we show that OmpF selectivity (traditionally regarded as slightly cationic) depends strongly on pH and salt concentration and is conditionally asymmetric, that is, the calculated selectivity is sensitive to the direction of salt concentration gradient. At neutral pH and subdecimolar salt concentrations the channel exhibits nearly ideal cation selectivity (t(G)(+)=0.98+/-0.01). Substituting neutral DPhPC with DPhPS, we demonstrate that the fixed charge of the host lipid has a small but measurable effect on the channel reversal potential. The available structural information allows for a qualitative explanation of our experimental findings. These findings now lead us to re-examine the ionization state of 102 titratable sites of the OmpF channel. Using standard methods of continuum electrostatics tailored to our particular purpose, we find the charge distribution in the channel as a function of solution acidity and relate the pH-dependent asymmetry in channel selectivity to the pH-dependent asymmetry in charge distribution. In an attempt to find a simple phenomenological description of our results, we also discuss different macroscopic models of electrodiffusion through large channels.

Anisotropy↗