8WA5

Cryo-EM structure of the gastric proton pump Y799W/E936Q mutant in K+-occluded (K+)E2-AlF state

8WA5 の概要

• エントリーDOI: 10.2210/pdb8wa5/pdb
• EMDBエントリー: 37391
• 分子名称: Sodium/potassium-transporting ATPase subunit alpha, Potassium-transporting ATPase subunit beta, POTASSIUM ION, ... (9 entities in total)
• 機能のキーワード: p-type atpase, gastric proton pump, membrane protein, primary transporter, transporter
• 由来する生物種: Sus scrofa (pig); 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 149920.83

構造登録者

Abe, K. (登録日: 2023-09-07, 公開日: 2023-11-29, 最終更新日: 2024-11-06)

主引用文献

Madapally, H.V.,Abe, K.,Dubey, V.,Khandelia, H.
Specific protonation of acidic residues confers K + selectivity to the gastric proton pump.
J.Biol.Chem., 300:105542-105542, 2023

PubMed Abstract: The gastric proton pump (H,K-ATPase) transports a proton into the stomach lumen for every K ion exchanged in the opposite direction. In the lumen-facing state of the pump (E2), the pump selectively binds K despite the presence of a 10-fold higher concentration of Na. The molecular basis for the ion selectivity of the pump is unknown. Using molecular dynamics simulations, free energy calculations, and Na and K-dependent ATPase activity assays, we demonstrate that the K selectivity of the pump depends upon the simultaneous protonation of the acidic residues E343 and E795 in the ion-binding site. We also show that when E936 is protonated, the pump becomes Na sensitive. The protonation-mimetic mutant E936Q exhibits weak Na-activated ATPase activity. A 2.5-Å resolution cryo-EM structure of the E936Q mutant in the K-occluded E2-Pi form shows, however, no significant structural difference compared with wildtype except less-than-ideal coordination of K in the mutant. The selectivity toward a specific ion correlates with a more rigid and less fluctuating ion-binding site. Despite being exposed to a pH of 1, the fundamental principle driving the K ion selectivity of H,K-ATPase is similar to that of Na,K-ATPase: the ionization states of the acidic residues in the ion-binding sites determine ion selectivity. Unlike the Na,K-ATPase, however, protonation of an ion-binding glutamate residue (E936) confers Na sensitivity.

PubMed: 38072058
DOI: 10.1016/j.jbc.2023.105542

実験手法

ELECTRON MICROSCOPY (2.51 Å)