Files
Abstract
Excitatory amino acid transporters (EAATs) maintain low extracellular glutamate levels and prevent excitotoxicity, and their dysfunction is linked to several neurological diseases. This work examines structural and functional properties of glutamate transporters using double site‑directed spin labeling electron paramagnetic resonance (DSDSL‑EPR) on the bacterial homolog GltPh. By measuring distance changes between spin labels, we monitored conformational shifts during ion, substrate, and inhibitor binding. Our findings validate electrophysiological and structural models of extracellular gating and reveal previously undescribed conformational changes in the apo‑state triggered by ion binding, providing new insight into transporter gating mechanisms relevant to therapeutic development.