Adrian H Elcock, Razif R Gabdoulline, Rebecca C Wade, J.Andrew McCammon

Computer simulations were performed to investigate the role of electrostatic interactions in promoting fast association of acetylcholinesterase with its peptidic inhibitor, the neurotoxin fasciculin. The encounter of the two macromolecules was simulated with the technique of Brownian dynamics (BD), using atomically detailed structures, and association rate constants were calculated for the wild-type and a number of mutant proteins. In a first set of simulations, the ordering of the experimental rate constants for the mutant proteins was correctly reproduced, although the absolute values of the rate constants were overestimated by a factor of around 30. Rigorous calculations of the full electrostatic interaction energy between the two proteins indicate that this overestimation of association rates results at least in part from approximations made in the description of interaction energetics in the BD simulations. In particular, the initial BD simulations neglect the unfavourable electrostatic desolvation effects that result from the exclusion of high dielectric solvent that accompanies the approach of the two low dielectric proteins. This electrostatic desolvation component is so large that the overall contribution of electrostatics to the binding energy of the complex is unlikely to be strongly favourable. Nevertheless, electrostatic interactions are still responsible for increased association rates, because even if they are unfavourable in the fully formed complex, they are still favourable at intermediate protein-protein separation distances. It therefore appears possible for electrostatic interactions to promote the kinetics of binding even if they do not make a strongly favourable contribution to the thermodynamics of binding. When an approximate description of these electrostatic desolvation effects is included in a second set of BD simulations, the relative ordering of the mutant proteins is again correctly reproduced, but now association rate constants that are much closer in magnitude to the experimental values are obtained. Inclusion of electrostatic desolvation effects also improves reproduction of the experimental ionic strength dependence of the wild-type association rate.