Charge Discrimination in P2X₄ Receptors Occurs in Two Consecutive Stages

P2X receptors are a group of trimeric cationic channels that are activated by adenosine 5'-triphosphate. They perform critical roles in the membranes of mammalian cells, and their improper functioning is associated with numerous diseases. Despite the vast amount of research devoted to them, several aspects of their operation are currently unclear, including the causes of their charge selectivity. We present the results of molecular dynamics simulation, which shed light on this issue for the case of P2X₄ channels. We examined in detail the behavior of Na⁺ and Cl^- ions inside the receptor. The examination reveals that charge discrimination occurs in two stages. First, cations bear precedence over anions to enter the extracellular vestibule. Then, cations at the extracellular vestibule are more likely to cross the pore than anions in an equivalent position. In this manner, a thorough but straightforward analysis of computational simulations suggests a stepwise mechanism, without a unique determinant factor.