Translocation Mechanism of Allosteric Sodium Ions in β₂-Adrenoceptor

The allosteric modulation of G-protein-coupled receptors (GPCRs) by sodium ions has received significant attention as the crystal structures of several receptors show the binding of sodium ions (Na⁺) at the conserved D^2.50. Theoretical studies have shown that extracellular Na⁺ would enter the allosteric D^2.50 via the orthosteric site. However, it remains unclear how the bound allosteric Na⁺ would leave the GPCRs. In this study, we performed molecular dynamics (MD) simulations to illustrate the energy barriers of Na⁺ transfer through the transmembrane helix bundle of β₂AR. In contrast to the postulations from other GPCRs, the translocation of this allosteric Na⁺ into the intracellular side is found to be significantly difficult. Hence, the translocation direction could be receptor-specific.