2026-04-13T16:52:58Zhttps://recercat.cat/oai/request

oai:recercat.cat:2445/2174492025-11-19T19:20:16Zcom_2072_1057col_2072_478781col_2072_478908col_2072_478917

00925njm 22002777a 4500 dc Fort i Baixeras, Joana author Nicolas Arago, Adria author Maggi, Luca author Martinez Molledo, Maria author Kapiki, Despoina author Gonzalez Novoa, Paula author Gomez Gejo, Patricia author Zijlstra, Niels author Bodoy i Salvans, Susanna author Pardon, Els author Steyaert, Jan author Llorca, Oscar author Orozco López, Modesto author Cordes, Thorben author Palacín Prieto, Manuel author 2025-01-14T11:10:44Z 2025-01-14T11:10:44Z 2025-01-02 2025-01-13T10:53:48Z L-Amino acid transporters (LATs) play a key role in a wide range of physiological processes. Defects in LATs can lead to neurological disorders and aminoacidurias, while the overexpression of these transporters is related to cancer. BasC is a bacterial LAT transporter with an APC fold. In this study, to monitor the cytoplasmic motion of BasC, we developed a single-molecule Förster resonance energy transfer assay that can characterize the conformational states of the intracellular gate in solution at room temperature. Based on combined biochemical and biophysical data and molecular dynamics simulations, we propose a model in which the conserved lysine residue in TM5 supports TM1a to explore both open and closed states within the cytoplasmic gate under apo conditions. This equilibrium can be altered by substrates, mutation of conserved lysine 154 in TM5, or a transport-blocking nanobody interacting with TM1a. Overall, these findings provide insights into the transport mechanism of BasC and highlight the significance of the lysine residue in TM5 in the cytoplasmic gating of LATs. Aminoàcids Transport biològic Anàlisi conformacional Amino acids Biological transport Conformational analysis The conserved lysine residue in transmembrane helix 5 is key for the cytoplasmic gating of the L-amino acid transporters