Ação de anestésicos gerais em canais iônicos

General anesthetics are daily used in a myriad of surgical procedures worldwide. Nevertheless their molecular mechanism of action remains elusive. An accredited hypothesis suggests that anesthetics modulate distinct ion channels in neural tissue by interacting in multiple binding sites in these membrane proteins. New in vitro studies show that halogenated anesthetics modifies the conductance/voltage relation of voltage gated potassium channels by binding to multiple cavities with different affinities in a conformationdependent fashion. Within this context, the present investigation is focused not only on searching the putative binding sites, but also on discovering the manner by which the binding could modify the conformation energy landscape, since the binding of the anesthetic can be a necessary but not sufficient condition to have a modulatory effect. The mammal Kv1.2 channel stands as a potential target of inhalation anesthetics. Sevoflurane, a halogenated anesthetic, potentiates this channel, shifting the conductance/voltage relation leftward and increasing the maximum conductance. Recently, the G329 point mutation in this channel has been associated with an augmentation of the sensitivity to the general anesthetic sevoflurane. To explore this issue, molecular dynamics simulations of the Kv1.2 wild type and Kv1.2 G329T inserted in a phospholipidic membrane with explicit water treatment were performed. The sampling was performed to both open and closed conformations. A slightly reduction of fluctuation near the linker S4-S5 was noticed on the mutant by analysis of RMSF through the simulated trajectory. From these simulations, 120 frames were selected from each construction for running docking calculation. These approach was used in order to account for molecular flexibility of the protein. From the molecular docking applying AutoDock Vina, nearly 2400 binding modes were obtained. After clustering and analysis of solutions, four putative independent binding cavities were selected to construct the anesthetic/channel complex and then ligand site-specific affinities were estimated by free energy calculations based on molecular dynamics simulations. By employing the LIE (Linear Interaction Energy) method, a relative strong affinity by the site near the selectivity filter was noted. The interaction with this site could favor the stabilization of the conductive state, hence increasing the conductance of the channel. Since the G329T mutation is associated with an increased equilibrium constant of the