6QV0

Structure of ATP-bound outward-facing TM287/288 in complex with sybody Sb_TM35

6QV0 の概要

• エントリーDOI: 10.2210/pdb6qv0/pdb
• 分子名称: ABC transporter, ATP-binding protein, Uncharacterized ABC transporter ATP-binding protein TM_0288, Sb_TM35, ... (5 entities in total)
• 機能のキーワード: abc exporter, abc transporter, membrane transporter, membrane protein, sybody, nanobody
• 由来する生物種: Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099); 詳細
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 295363.70

構造登録者

Hutter, C.A.J.,Huerlimann, L.M.,Zimmermann, I.,Egloff, P.,Seeger, M.A. (登録日: 2019-03-01, 公開日: 2019-05-29, 最終更新日: 2024-11-20)

主引用文献

Hutter, C.A.J.,Timachi, M.H.,Hurlimann, L.M.,Zimmermann, I.,Egloff, P.,Goddeke, H.,Kucher, S.,Stefanic, S.,Karttunen, M.,Schafer, L.V.,Bordignon, E.,Seeger, M.A.
The extracellular gate shapes the energy profile of an ABC exporter.
Nat Commun, 10:2260-2260, 2019

PubMed Abstract: ABC exporters harness the energy of ATP to pump substrates across membranes. Extracellular gate opening and closure are key steps of the transport cycle, but the underlying mechanism is poorly understood. Here, we generated a synthetic single domain antibody (sybody) that recognizes the heterodimeric ABC exporter TM287/288 exclusively in the presence of ATP, which was essential to solve a 3.2 Å crystal structure of the outward-facing transporter. The sybody binds to an extracellular wing and strongly inhibits ATPase activity by shifting the transporter's conformational equilibrium towards the outward-facing state, as shown by double electron-electron resonance (DEER). Mutations that facilitate extracellular gate opening result in a comparable equilibrium shift and strongly reduce ATPase activity and drug transport. Using the sybody as conformational probe, we demonstrate that efficient extracellular gate closure is required to dissociate the NBD dimer after ATP hydrolysis to reset the transporter back to its inward-facing state.

PubMed: 31113958
DOI: 10.1038/s41467-019-09892-6

実験手法

X-RAY DIFFRACTION (3.12 Å)