Ionic strength dependence of F-actin and glycolytic enzyme associations: a Brownian dynamics simulations approach.

The association of glycolytic enzymes with F-actin is proposed to be one mechanism by which these enzymes are compartmentalized, and, as a result, may possibly play important roles for: regulation of the glycolytic pathway, potential substrate channeling, and increasing glycolytic flux. Historically, in vitro experiments have shown that many enzyme/actin interactions are dependent on ionic strength. Herein, Brownian dynamics (BD) examines how ionic strength impacts the energetics of the association of F-actin with the glycolytic enzymes: lactate dehydrogenase (LDH), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), fructose-1,6-bisphosphate aldolase (aldolase), and triose phosphate isomerase (TPI). The BD simulations are steered by electrostatics calculated by Poisson-Boltzmann theory. The BD results confirm experimental observations that the degree of association diminishes as ionic strength increases but also suggest that these interactions are significant, at physiological ionic strengths. Furthermore, BD agrees with experiments that muscle LDH, aldolase, and GAPDH interact significantly with F-actin whereas TPI does not. BD indicates similarities in binding regions for aldolase and LDH among the different species investigated. Furthermore, the residues responsible for salt bridge formation in stable complexes persist as ionic strength increases. This suggests the importance of the residues determined for these binary complexes and specificity of the interactions. That these interactions are conserved across species, and there appears to be a general trend among the enzymes, support the importance of these enzyme-F-actin interactions in creating initial complexes critical for compartmentation.

BD SIMULATIONS OF THE IONIC STRENGTH DEPENDENCE OF THE INTERACTIONS BETWEEN TRIOSE PHOSPHATE ISOMERASE AND F-ACTIN.

Functional protein-protein interactions are essential for many physiological processes. For example, the association of glycolytic enzymes to F-actin is proposed to be one mechanism through which glycolytic enzymes are compartmentalized, and as a result, play essential roles such as regulation of the glycolytic pathway and increasing glycolytic flux. Many glycolytic enzymes including fructose-1,6-bisphophate aldolase, glyceraldedhye-3-phosphate dehydrogenase, and lactate dehydrogenase bind F-actin strongly. Other glycolytic enzymes including triose phosphate isomerase (TPI) do not interact with F-actin significantly. Herein, Brownian dynamics (BD) simulations determine the energetics of the association of F-actin with the glycolytic enzyme triose phosphate isomerase as a function of ionic strength. This is the first thorough control study examining how well BD reproduces the experimental observations that the binding of TPI to F-actin is very weak and falls off rapidly as ionic strength increases. The BD results confirm experimental observations that the degree of association diminishes as ionic strength increases and that the interaction of TPI with F-actin is weakly nonspecific to nonexistent.