8PDA

cryo-EM structure of Doa10 with RING domain in MSP1E3D1

8PDA の概要

• エントリーDOI: 10.2210/pdb8pda/pdb
• EMDBエントリー: 17608
• 分子名称: ERAD-associated E3 ubiquitin-protein ligase DOA10, 1,2-DIOLEOYL-SN-GLYCERO-3-PHOSPHOCHOLINE, 1,2-DIPALMITOYL-SN-GLYCERO-3-PHOSPHATE (3 entities in total)
• 機能のキーワード: erad, doa10, march6, teb4, retrotranslocation, ubiquitination, ubc6, sybody, sqle, squalenemonooxygenase, ligase
• 由来する生物種: Saccharomyces cerevisiae (baker's yeast)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 154617.36

構造登録者

Botsch, J.J.,Braeuning, B.,Schulman, B.A. (登録日: 2023-06-12, 公開日: 2024-01-17)

主引用文献

Botsch, J.J.,Junker, R.,Sorgenfrei, M.,Ogger, P.P.,Stier, L.,von Gronau, S.,Murray, P.J.,Seeger, M.A.,Schulman, B.A.,Brauning, B.
Doa10/MARCH6 architecture interconnects E3 ligase activity with lipid-binding transmembrane channel to regulate SQLE.
Nat Commun, 15:410-410, 2024

PubMed Abstract: Transmembrane E3 ligases play crucial roles in homeostasis. Much protein and organelle quality control, and metabolic regulation, are determined by ER-resident MARCH6 E3 ligases, including Doa10 in yeast. Here, we present Doa10/MARCH6 structural analysis by cryo-EM and AlphaFold predictions, and a structure-based mutagenesis campaign. The majority of Doa10/MARCH6 adopts a unique circular structure within the membrane. This channel is established by a lipid-binding scaffold, and gated by a flexible helical bundle. The ubiquitylation active site is positioned over the channel by connections between the cytosolic E3 ligase RING domain and the membrane-spanning scaffold and gate. Here, by assaying 95 MARCH6 variants for effects on stability of the well-characterized substrate SQLE, which regulates cholesterol levels, we reveal crucial roles of the gated channel and RING domain consistent with AlphaFold-models of substrate-engaged and ubiquitylation complexes. SQLE degradation further depends on connections between the channel and RING domain, and lipid binding sites, revealing how interconnected Doa10/MARCH6 elements could orchestrate metabolic signals, substrate binding, and E3 ligase activity.

PubMed: 38195637
DOI: 10.1038/s41467-023-44670-5

実験手法

ELECTRON MICROSCOPY (3.58 Å)