6SI7

Structure of the curli secretion-assembly complex CsgG:CsgF

6SI7 の概要

• エントリーDOI: 10.2210/pdb6si7/pdb
• EMDBエントリー: 10206
• 分子名称: Curli production assembly/transport component CsgF, Curli production assembly/transport component CsgG (2 entities in total)
• 機能のキーワード: secretion channel, curli, outer membrane protein, nanopore sensing, protein transport, bacterial amyloid
• 由来する生物種: Escherichia coli; 詳細
• タンパク質・核酸の鎖数: 18
• 化学式量合計: 394695.45

構造登録者

Van der Verren, S.E.,Remaut, H. (登録日: 2019-08-08, 公開日: 2020-06-24, 最終更新日: 2024-05-22)

主引用文献

Van der Verren, S.E.,Van Gerven, N.,Jonckheere, W.,Hambley, R.,Singh, P.,Kilgour, J.,Jordan, M.,Wallace, E.J.,Jayasinghe, L.,Remaut, H.
A dual-constriction biological nanopore resolves homonucleotide sequences with high fidelity.
Nat.Biotechnol., 38:1415-1420, 2020

PubMed Abstract: Single-molecule long-read DNA sequencing with biological nanopores is fast and high-throughput but suffers reduced accuracy in homonucleotide stretches. We now combine the CsgG nanopore with the 35-residue N-terminal region of its extracellular interaction partner CsgF to produce a dual-constriction pore with improved signal and base-calling accuracy for homopolymer regions. The electron cryo-microscopy structure of CsgG in complex with full-length CsgF shows that the 33 N-terminal residues of CsgF bind inside the β-barrel of the pore, forming a defined second constriction. In complexes of CsgG bound to a 35-residue CsgF constriction peptide, the second constriction is separated from the primary constriction by ~25 Å. We find that both constrictions contribute to electrical signal modulation during single-stranded DNA translocation. DNA sequencing using a prototype CsgG-CsgF protein pore with two constrictions improved single-read accuracy by 25 to 70% in homopolymers up to 9 nucleotides long.

PubMed: 32632300
DOI: 10.1038/s41587-020-0570-8

実験手法

ELECTRON MICROSCOPY (3.4 Å)