9H76

Bacterial LAT transporter BASC in complex with L-Ala and NB53

Summary for 9H76

• Entry DOI: 10.2210/pdb9h76/pdb
• Related: 6F2G 6F2W
• EMDB information: 51912
• Descriptor: Putative amino acid/polyamine transport protein, Nanobody NB53, ALANINE (3 entities in total)
• Functional Keywords: transporter, nanobody, aminoacid transporter, membrane protein
• Biological source: Carnobacterium sp. AT7; More
• Total number of polymer chains: 2
• Total formula weight: 61043.49

Authors

Fort, J.,Palacin, M. (deposition date: 2024-10-26, release date: 2025-01-01, Last modification date: 2025-07-02)

Primary citation

Fort, J.,Nicolas-Arago, A.,Maggi, L.,Martinez-Molledo, M.,Kapiki, D.,Gonzalez-Novoa, P.,Gomez-Gejo, P.,Zijlstra, N.,Bodoy, S.,Pardon, E.,Steyaert, J.,Llorca, O.,Orozco, M.,Cordes, T.,Palacin, M.
The conserved lysine residue in transmembrane helix 5 is pivotal for the cytoplasmic gating of the L-amino acid transporters.
Pnas Nexus, 4:pgae584-pgae584, 2025

PubMed Abstract: 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.

PubMed: 39822574
DOI: 10.1093/pnasnexus/pgae584

Experimental method

ELECTRON MICROSCOPY (2.9 Å)