Publication

Preparation of Fragaceatoxin C (FraC) Nanopores

Mutter, N. L., Huang, G., van der Heide, N. J., Lucas, F. L. R., Galenkamp, N. S., Maglia, G. & Wloka, C., 2021, Nanopore Technology: Methods in Molecular Biology. Fahie, M. A. (ed.). New York: Humana Press, p. 3-10 8 p. (Methods in Molecular Biology; vol. 2186).

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

Biological nanopores are an emerging class of biosensors with high-end precision owing to their reproducible fabrication at the nanometer scale. Most notably, nanopore-based DNA sequencing applications are currently being commercialized, while nanopore-based proteomics may become a reality in the near future.Although membrane proteins often prove to be difficult to purify, we describe a straightforward protocol for the preparation of Fragaceatoxin C (FraC) nanopores, which may have applications for DNA analysis and nanopore-based proteomics. Recombinantly expressed FraC nanopores are purified via two rounds of Ni-NTA affinity chromatography before and after oligomerization on sphingomyelin-containing liposomes. Starting from a plasmid vector containing the FraC gene, our method allows the production of purified nanopores within a week. Afterward, the FraC nanopores can be stored at +4 °C for several months, or frozen.

Original languageEnglish
Title of host publicationNanopore Technology
Subtitle of host publicationMethods in Molecular Biology
EditorsM.A. Fahie
Place of PublicationNew York
PublisherHumana Press
Pages3-10
Number of pages8
ISBN (Electronic)978-1-0716-0806-7
ISBN (Print)978-1-0716-0805-0
Publication statusPublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2186
ISSN (Print)1064-3745

    Keywords

  • Nanotechnology, Nanopore, Porin, actinoporin, Fragaceatoxin C, Fra C, protein purification, protein oligomerization, electrophysiology, artificial bilayers

ID: 133662779