7NGH

Structure of glutamate transporter homologue in complex with Sybody

7NGH の概要

• エントリーDOI: 10.2210/pdb7ngh/pdb
• 関連するPDBエントリー: 6XWQ
• EMDBエントリー: 12314
• 分子名称: Proton/glutamate symporter, SDF family, Sybody 1, ASPARTIC ACID (3 entities in total)
• 機能のキーワード: glutamate transporter homologue, glttk, amino acid transport, membrane protein, transport protein
• 由来する生物種: Thermococcus kodakarensis KOD1; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 155488.92

構造登録者

Arkhipova, V.,Slotboom, D.J.,Guskov, A. (登録日: 2021-02-09, 公開日: 2021-09-15, 最終更新日: 2025-07-09)

主引用文献

Trinco, G.,Arkhipova, V.,Garaeva, A.A.,Hutter, C.A.J.,Seeger, M.A.,Guskov, A.,Slotboom, D.J.
Kinetic mechanism of Na + -coupled aspartate transport catalyzed by Glt Tk .
Commun Biol, 4:751-751, 2021

PubMed Abstract: It is well-established that the secondary active transporters Glt and Glt catalyze coupled uptake of aspartate and three sodium ions, but insight in the kinetic mechanism of transport is fragmentary. Here, we systematically measured aspartate uptake rates in proteoliposomes containing purified Glt, and derived the rate equation for a mechanism in which two sodium ions bind before and another after aspartate. Re-analysis of existing data on Glt using this equation allowed for determination of the turnover number (0.14 s), without the need for error-prone protein quantification. To overcome the complication that purified transporters may adopt right-side-out or inside-out membrane orientations upon reconstitution, thereby confounding the kinetic analysis, we employed a rapid method using synthetic nanobodies to inactivate one population. Oppositely oriented Glt proteins showed the same transport kinetics, consistent with the use of an identical gating element on both sides of the membrane. Our work underlines the value of bona fide transport experiments to reveal mechanistic features of Na-aspartate symport that cannot be observed in detergent solution. Combined with previous pre-equilibrium binding studies, a full kinetic mechanism of structurally characterized aspartate transporters of the SLC1A family is now emerging.

PubMed: 34140623
DOI: 10.1038/s42003-021-02267-y

実験手法

ELECTRON MICROSCOPY (3.5 Å)