Mechanism of μ-Opioid Receptor-Magnesium Interaction and Positive Allosteric Modulation.

Abstract

In the era of opioid abuse epidemics, there is an increased demand for understanding how opioid receptors can be allosterically modulated to guide the development of more effective and safer opioid therapies. Among the modulators of the {micro}-opioid (MOP) receptor, which is the pharmacological target for the majority of clinically used opioid drugs, are monovalent and divalent cations. Specifically, the monovalent sodium cation (Na+) has been known for decades to affect MOP receptor signaling by reducing agonist binding, whereas the divalent magnesium cation (Mg2+) has been shown to have the opposite effect, notwithstanding the presence of sodium chloride. While ultra-high resolution opioid receptor crystal structures have revealed a specific Na+ binding site and molecular dynamics (MD) simulation studies have supported the idea that this monovalent ion reduces agonist binding by stabilizing the receptor inactive state, the putative binding site of Mg2+ on the MOP receptor, as well as the molecular determinants responsible for its positive allosteric modulation of the receptor, are unknown. In this work, we carried out tens of microseconds of all-atom MD simulations to investigate the simultaneous binding of Mg2+ and Na+ cations to inactive and active crystal structures of the MOP receptor embedded in an explicit lipid/water environment. Analyses of these simulations shed light on (a) the preferred binding sites of Mg2+ on the MOP receptor, (b) details of the competition between Mg2+ and Na+ cations for specific sites, (c) estimates of binding affinities, and (d) testable hypotheses of the molecular mechanism underlying the positive allosteric modulation of the MOP receptor by the Mg2+ cation.\n\nStatement of SignificanceOverprescription of opioid drugs in the late 1990s, followed by abuse of both prescription and illicit opioid drugs, has led to what is nowadays called \"opioid overdose crisis\" or \"opioid epidemic\", with more than 130 Americans dying daily from opioid overdose at the time of writing. To reduce opioid doses, and thereby prevent or treat overdose and opioid use disorders, attention has recently shifted to the use of co-analgesics. Understanding how opioid receptor targets can be allosterically modulated by these elements, including cations, is key to the development of improved therapeutics. Here, we provide an atomic-level understanding of the mechanism by which magnesium binds to the {micro}-opioid receptor and enhances opioid drug efficacy by stabilizing the receptor activated state.