Preparation of Fragaceatoxin C (FraC) Nanopores

Natalie Lisa Mutter, Gang Huang, Nieck Jordy van der Heide, Florian Leonardus Rudolfus Lucas, Nicole Stéphanie Galenkamp, Giovanni Maglia, Carsten Wloka*

*Corresponding author for this work

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

4 Citations (Scopus)


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
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
ISSN (Print)1064-3745


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

Cite this