EMDB/PDB/SASBDBから引用されている文献のデータベース

キーワード
構造解析手法	All X線回折 NMR 電子顕微鏡小角散乱中性子線回折線維回折粉末回折赤外分光 EPR FRET 理論モデルハイブリッド
著者・登録者
ジャーナル
IF	>30 >20 >10 >5 all

タイトル	Structural basis of the excitatory amino acid transporter 3 substrate recognition.
ジャーナル・号・ページ	bioRxiv, Year 2024
掲載日	2024年9月8日
著者	Biao Qiu / Olga Boudker /
PubMed 要旨	Excitatory amino acid transporters (EAATs) reside on cell surfaces and uptake substrates, including L-glutamate, L-aspartate, and D-aspartate, using ion gradients. Among five EAATs, EAAT3 is the only ...Excitatory amino acid transporters (EAATs) reside on cell surfaces and uptake substrates, including L-glutamate, L-aspartate, and D-aspartate, using ion gradients. Among five EAATs, EAAT3 is the only isoform that can efficiently transport L-cysteine, a substrate for glutathione synthesis. Recent work suggests that EAAT3 also transports the oncometabolite R-2-hydroxyglutarate (R-2HG). Here, we examined the structural basis of substrate promiscuity by determining the cryo-EM structures of EAAT3 bound to different substrates. We found that L-cysteine binds to EAAT3 in thiolate form, and EAAT3 recognizes different substrates by fine-tuning local conformations of the coordinating residues. However, using purified human EAAT3, we could not observe R-2HG binding or transport. Imaging of EAAT3 bound to L-cysteine revealed several conformational states, including an outward-facing state with a semi-open gate and a disrupted sodium-binding site. These structures illustrate that the full gate closure, coupled with the binding of the last sodium ion, occurs after substrate binding. Furthermore, we observed that different substrates affect how the transporter distributes between a fully outward-facing conformation and intermediate occluded states on a path to the inward-facing conformation, suggesting that translocation rates are substrate-dependent.
リンク	bioRxiv / PubMed:39282329 / PubMed Central
手法	EM (単粒子)
解像度	2.58 - 3.07 Å
構造データ	46586 9d66 EMDB-46586, PDB-9d66: Human excitatory amino acid transporter 3 (EAAT3) with bound L-Aspartate in an intermediate outward facing state 手法: EM (単粒子) / 解像度: 2.98 Å EMDB-46587: EAAT3 in 200 mM NaCl and 10 mM R-2-hydroxyglutarate buffer at OFS. 手法: EM (単粒子) / 解像度: 3.07 Å 46588 9d67 EMDB-46588, PDB-9d67: Human excitatory amino acid transporter 3 (EAAT3) with bound D-Aspartate in an intermediate outward facing state 手法: EM (単粒子) / 解像度: 2.87 Å 46589 9d68 EMDB-46589, PDB-9d68: Human excitatory amino acid transporter 3 (EAAT3) with bound L-Cysteine in an outward facing state 手法: EM (単粒子) / 解像度: 2.58 Å 46590 9d69 EMDB-46590, PDB-9d69: Human excitatory amino acid transporter 3 (EAAT3) with bound L-Cysteine in an intermediate outward facing state (slight upper position) 手法: EM (単粒子) / 解像度: 2.99 Å 46591 9d6a EMDB-46591, PDB-9d6a: Human excitatory amino acid transporter 3 (EAAT3) with bound L-Cysteine in an intermediate outward facing state 手法: EM (単粒子) / 解像度: 2.6 Å EMDB-46592: Human excitatory amino acid transporter 3 (EAAT3) monomer in 200 mM NaCl, 10 mM L-Cysteine at IFS 手法: EM (単粒子) / 解像度: 2.94 Å
化合物	ChemComp-ASP: ASPARTIC ACID / アスパラギン酸 ChemComp-NA: Unknown entry / ナトリウムカチオン ChemComp-HG: Unknown entry ChemComp-HOH: WATER ChemComp-DAS: D-ASPARTIC ACID / D-アスパラギン酸 ChemComp-CYS: CYSTEINE / システイン
由来	homo sapiens (ヒト)
キーワード	TRANSPORT PROTEIN / EAAT3 / L-Asp / iOFS / human EAAT3 / D-Asp / Cysteine / OFS / semi-open / iOFS-upper / PROTEIN TRANSPORT / iOFS-star