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| Title | A dual-constriction biological nanopore resolves homonucleotide sequences with high fidelity. |
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| Journal, issue, pages | Nat Biotechnol, Vol. 38, Issue 12, Page 1415-1420, Year 2020 |
| Publish date | Jul 6, 2020 |
 Authors | Sander E Van der Verren / Nani Van Gerven / Wim Jonckheere / Richard Hambley / Pratik Singh / John Kilgour / Michael Jordan / E Jayne Wallace / Lakmal Jayasinghe / Han Remaut /   |
| 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- ...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. |
 External links | Nat Biotechnol / PubMed:32632300 / PubMed Central |
| Methods | EM (single particle) |
| Resolution | 3.4 Å |
| Structure data | EMDB-10206, PDB-6si7: |
| Source |
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 Keywords | PROTEIN TRANSPORT / Secretion Channel / Curli / Outer Membrane Protein / Nanopore Sensing / Bacterial amyloid |
escherichia coli (E. coli)
