Impact of anionic lipids on the energy landscape of conformational transition in anion exchanger 1 (AE1).

Chen, Tianle; Vallese, Francesca; Gil-Iturbe, Eva; Kim, Kookjoo; Calì, Tito; Quick, Matthias; Clarke, Oliver B; Tajkhorshid, Emad · Nat Commun · 2025

basic_science · Level V

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Abstract

Anion Exchanger 1 (AE1) is an elevator-type transporter that plays a key role in acid-base homeostasis of erythrocytes. Here, we report three high-resolution cryo-electron microscopy (cryo-EM) structures of distinct states of AE1: two inward-facing (IF1 and IF2) and one outward-facing (OF). Uptake assay revealed the modulatory effect of phosphatidylinositol 4,5-bisphosphate (PIP<sub>2</sub>) lipids on AE1. Molecular dynamics simulations are conducted on these structures to determine the anion binding sites in AE1. We then use advanced enhanced sampling to study the OF⇌IF transition in AE1 in three systems: apo, HCO<sub>3</sub><sup>-</sup>-bound, and an AE1 system in which cryo-EM-determined PIP<sub>2</sub> lipids had been removed. The transition pathways were then used to calculate the free energy of the OF⇌IF transition in AE1 under different conditions. The results show how substrate reduces the transition barrier against transport. Furthermore, they capture the inhibitory effect of PIP<sub>2</sub> lipids and provide a molecular mechanism for this inhibitory effect.

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