4.7 Article

Synthesis, lipid membrane incorporation, and ion permeability testing of carbon nanotube porins

Journal

NATURE PROTOCOLS
Volume 11, Issue 10, Pages 2029-2047

Publisher

NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2016.119

Keywords

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Funding

  1. US Department of Energy, Office of Basic Energy Sciences, Division of Materials Sciences and Engineering
  2. LDRD program at LLNL [12-ERD-073]
  3. US Department of Energy [DE-AC52-07NA27344]
  4. Office of Science, Office of Basic Energy Sciences, of the US Department of Energy [DE-AC02-05CH11231]
  5. Spanish Ministry of Economy and Competitiveness [BIO2013-49843-EXP]

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Carbon nanotube porins (CNTPs) are 10- to 20-nm-long segments of lipid-stabilized single-walled carbon nanotubes (CNTs) that can be inserted into phospholipid membranes to form nanometer-scale-diameter pores that approximate the geometry and many key transport characteristics of biological membrane channels. We describe protocols for CNTP synthesis by ultrasound-assisted cutting of long CNTs in the presence of lipid amphiphiles, and for validation of CNTP incorporation into a lipid membrane using a proton permeability assay. In addition, we describe protocols for measuring conductance of individual CNTPs in planar lipid bilayers and plasma membranes of live cells. The protocol for the preparation and testing of the CNTPs in vesicle systems takes 3 d, and single CNTP conductance measurements take 2-5 h. The CNTPs produced by this cutting protocol remain stable and active for at least 10-12 weeks.

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