Role of the lipid matrix in the action of local anesthetics

The binding of local anesthetics (LAs) to cell membranes is required for LAs to reach the target ion channels, but lipid interactions may also play a role in a purely membrane-mediated mode of activity. Here, we used solid-state NMR and further biophysical techniques to characterize the effect of six LAs covering a wide range of structures and properties (benzocaine, bupivacaine, mepivacaine, lidocaine, procaine, QX-314) on membranes. Membrane partitioning log D values (between 2.1 and 3.7) varied little with pH, in contrast to octanol partitioning. Membrane thinning was induced by most LAs, except for benzocaine. A conformational change in the lipid headgroup was observed, with a pronounced dependence on the protonation state, indicating the importance of the positive charge that is maintained by most membrane-bound LAs. We found stabilization of negative membrane curvature in the case of benzocaine, and of positive curvature in the case of bupivacaine, procaine, mepivacaine and, most pronouncedly, for QX-314. Comparing the LAs with respect of their influence on membranes as observed in the different experiments, benzocaine and QX-314 were always found at either extreme of the scale, with bupivacaine and lidocaine closer in their effect to benzocaine. This order of influence correlates with the depth of membrane insertion and with the protonation state, both of which were identified as key factors for LA behavior. Finally, we found indications that LAs are able to alter the activity of bacterial mechanosensitive channels without any expected LA binding sites, thus supporting a membrane-mediated activity of LAs.