Structural insights into a citrate transporter that mediates aluminum tolerance in barley

• 成果类型: Article
• 署名作者: Thao, Tran Nguyen; Mitani-Ueno, Namiki; Urano, Ryo; Saitoh, Yasunori; Wang, Peitong; Yamaji, Naoki; Shen, Jian-Ren; Shinoda, Wataru; Ma, Jian Feng; Suga, Michihiro
• 署名单位: Okayama University; Okayama University; Okayama University; Okayama University; Nanjing Agricultural University
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-12879
• DOI: 10.1073/pnas.2501933122
• 发表日期: 2025-08-05
• 关键词: gui membrane-builder; additive force-field; crystal-structure; multidrug; extrusion; acidification; ACID; translocation; sensitivity; resistance

摘要

HvAACT1 is a major aluminum (Al)-tolerance gene in barley, encoding a citrate transporter that belongs to the multidrug and toxic compound extrusion (MATE) family. This transporter facilitates citrate secretion from the roots, thereby detoxifying external Al ions-a major constraint of crop production on acidic soils. In this study, we present the outward-facing crystal structure of HvAACT1, providing insights into a citrate transport mechanism. The putative citrate binding site consists of three basic residues-K126 in transmembrane helix 2 (TM2), R358 in TM7, and R535 in TM12-creating substantial positive charges in the C-lobe cavity. Proton coupling for substrate transport may involve two pairs of aspartate residues in the N-lobe cavity, one of which corresponds to the essential Asp pair found in prokaryotic H+-coupled MATE transporters belonging to the DinF subfamily. Structural coupling between proton uptake in the N-lobe and citrate extrusion in the C-lobe can be enabled by an extensive, unique hydrogen-bonding network at the extracellular half of the N-lobe. Mutation-based functional analysis, structural comparisons, molecular dynamics simulation, and phylogenic analysis suggest an evolutionary link between citrate MATE transporters and the DinF MATE subfamily. Our findings provide a solid structural basis for citrate transport by HvAACT1 in barley and contribute to a broader understanding of citrate transporter structures in other plant species.