8DKX

Cryo-EM structure of cystinosin N288K mutant in a cytosol-open state at pH7.5

8DKX の概要

• エントリーDOI: 10.2210/pdb8dkx/pdb
• EMDBエントリー: 27493
• 分子名称: Fab 3H5 Heavy Chain, Fab 3H5 Kappa Chain, Isoform 2 of Cystinosin (3 entities in total)
• 機能のキーワード: cystine, transporter, lysosome, membrane protein, membrane protein-transport protein complex, membrane protein/transport protein
• 由来する生物種: Mus musculus; 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 98331.95

構造登録者

Schmiege, P.,Li, X. (登録日: 2022-07-06, 公開日: 2022-09-21, 最終更新日: 2024-11-06)

主引用文献

Guo, X.,Schmiege, P.,Assafa, T.E.,Wang, R.,Xu, Y.,Donnelly, L.,Fine, M.,Ni, X.,Jiang, J.,Millhauser, G.,Feng, L.,Li, X.
Structure and mechanism of human cystine exporter cystinosin.
Cell, 185:3739-3752.e18, 2022

PubMed Abstract: Lysosomal amino acid efflux by proton-driven transporters is essential for lysosomal homeostasis, amino acid recycling, mTOR signaling, and maintaining lysosomal pH. To unravel the mechanisms of these transporters, we focus on cystinosin, a prototypical lysosomal amino acid transporter that exports cystine to the cytosol, where its reduction to cysteine supplies this limiting amino acid for diverse fundamental processes and controlling nutrient adaptation. Cystinosin mutations cause cystinosis, a devastating lysosomal storage disease. Here, we present structures of human cystinosin in lumen-open, cytosol-open, and cystine-bound states, which uncover the cystine recognition mechanism and capture the key conformational states of the transport cycle. Our structures, along with functional studies and double electron-electron resonance spectroscopic investigations, reveal the molecular basis for the transporter's conformational transitions and protonation switch, show conformation-dependent Ragulator-Rag complex engagement, and demonstrate an unexpected activation mechanism. These findings provide molecular insights into lysosomal amino acid efflux and a potential therapeutic strategy.

PubMed: 36113465
DOI: 10.1016/j.cell.2022.08.020

実験手法

ELECTRON MICROSCOPY (3 Å)