7OCY

Enterococcus faecalis EfrCD in complex with a nanobody

7OCY の概要

• エントリーDOI: 10.2210/pdb7ocy/pdb
• EMDBエントリー: 12816
• 分子名称: ABC transporter ATP-binding protein, Nanobody (3 entities in total)
• 機能のキーワード: abc transporter, nanobody, enterococcus faecalis, membrane protein
• 由来する生物種: Enterococcus faecalis; 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 141837.16

構造登録者

Ehrenbolger, K.,Hutter, C.A.J.,Meier, G.,Seeger, M.A.,Barandun, J. (登録日: 2021-04-28, 公開日: 2022-05-18, 最終更新日: 2024-07-10)

主引用文献

Meier, G.,Thavarasah, S.,Ehrenbolger, K.,Hutter, C.A.J.,Hurlimann, L.M.,Barandun, J.,Seeger, M.A.
Deep mutational scan of a drug efflux pump reveals its structure-function landscape.
Nat.Chem.Biol., 19:440-450, 2023

PubMed Abstract: Drug efflux is a common resistance mechanism found in bacteria and cancer cells, but studies providing comprehensive functional insights are scarce. In this study, we performed deep mutational scanning (DMS) on the bacterial ABC transporter EfrCD to determine the drug efflux activity profile of more than 1,430 single variants. These systematic measurements revealed that the introduction of negative charges at different locations within the large substrate binding pocket results in strongly increased efflux activity toward positively charged ethidium, whereas additional aromatic residues did not display the same effect. Data analysis in the context of an inward-facing cryogenic electron microscopy structure of EfrCD uncovered a high-affinity binding site, which releases bound drugs through a peristaltic transport mechanism as the transporter transits to its outward-facing conformation. Finally, we identified substitutions resulting in rapid Hoechst influx without affecting the efflux activity for ethidium and daunorubicin. Hence, single mutations can convert EfrCD into a drug-specific ABC importer.

PubMed: 36443574
DOI: 10.1038/s41589-022-01205-1

実験手法

ELECTRON MICROSCOPY (4.25 Å)