Chasing the full free energy landscape of neuroreceptor/ligand unbinding by metadynamics simulations

Abstract

Predicting the complete free energy landscape associated with protein-ligand un-binding would greatly help designing drugs with highly optimized pharmacokinetics. Here we investigate the unbinding of the iperoxo agonist to its target human neuroreceptor M2, embedded in a neuronal membrane. By feeding out-of-equilibrium molecular simulations data in a classification analysis, we identify the few essential reaction coordinates of the process. The full landscape is then reconstructed using an exact enhanced sampling method, well-tempered metadynamics in its funnel variant. The calculations reproduce well the measured affinity, provide a rationale for mutagenesis data and show that the ligand can escape via two different routes. The allosteric modulator LY2119620 turns out to hamper both escapes routes, thus slowing down the unbinding process, as experimentally observed. This computationally affordable protocol is totally general and it can be easily applied to determine the full free energy landscape of membrane receptors/drug interactions. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=81 SRC="FIGDIR/small/544577v1_ufig1.gif" ALT="Figure 1"> View larger version (26K): org.highwire.dtl.DTLVardef@cfc6b2org.highwire.dtl.DTLVardef@fde50corg.highwire.dtl.DTLVardef@c8a893org.highwire.dtl.DTLVardef@1b36a12_HPS_FORMAT_FIGEXP M_FIG Graphical TOC Entry C_FIG