6UWL

GltPh in complex with L-aspartate and sodium ions in intermediate outward-facing state

6UWL の概要

• エントリーDOI: 10.2210/pdb6uwl/pdb
• EMDBエントリー: 20923
• 分子名称: Glutamate transporter homolog, ASPARTIC ACID, [(2~{R})-1-[2-azanylethoxy(oxidanyl)phosphoryl]oxy-3-hexadecanoyloxy-propan-2-yl] (~{Z})-octadec-9-enoate (3 entities in total)
• 機能のキーワード: aspartate transporter, transport protein
• 由来する生物種: Pyrococcus horikoshii
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 45521.06

構造登録者

Wang, X.,Boudker, O. (登録日: 2019-11-05, 公開日: 2020-04-22, 最終更新日: 2024-12-25)

主引用文献

Huang, Y.,Wang, X.,Lv, G.,Razavi, A.M.,Huysmans, G.H.M.,Weinstein, H.,Bracken, C.,Eliezer, D.,Boudker, O.
Use of paramagnetic 19 F NMR to monitor domain movement in a glutamate transporter homolog.
Nat.Chem.Biol., 16:1006-1012, 2020

PubMed Abstract: In proteins where conformational changes are functionally important, the number of accessible states and their dynamics are often difficult to establish. Here we describe a novel F-NMR spectroscopy approach to probe dynamics of large membrane proteins. We labeled a glutamate transporter homolog with a F probe via cysteine chemistry and with a Ni ion via chelation by a di-histidine motif. We used distance-dependent enhancement of the longitudinal relaxation of F nuclei by the paramagnetic metal to assign the observed resonances. We identified one inward- and two outward-facing states of the transporter, in which the substrate-binding site is near the extracellular and intracellular solutions, respectively. We then resolved the structure of the unanticipated second outward-facing state by cryo-EM. Finally, we showed that the rates of the conformational exchange are accessible from measurements of the metal-enhanced longitudinal relaxation of F nuclei.

PubMed: 32514183
DOI: 10.1038/s41589-020-0561-6

実験手法

ELECTRON MICROSCOPY (3.62 Å)