Letter | Published:

Stochastic transport through carbon nanotubes in lipid bilayers and live cell membranes

Nature volume 514, pages 612615 (30 October 2014) | Download Citation

Abstract

There is much interest in developing synthetic analogues of biological membrane channels1 with high efficiency and exquisite selectivity for transporting ions and molecules. Bottom-up2 and top-down3 methods can produce nanopores of a size comparable to that of endogenous protein channels, but replicating their affinity and transport properties remains challenging. In principle, carbon nanotubes (CNTs) should be an ideal membrane channel platform: they exhibit excellent transport properties4,5,6,7,8 and their narrow hydrophobic inner pores mimic structural motifs typical of biological channels1. Moreover, simulations predict that CNTs with a length comparable to the thickness of a lipid bilayer membrane can self-insert into the membrane9,10. Functionalized CNTs have indeed been found to penetrate lipid membranes and cell walls11,12, and short tubes have been forced into membranes to create sensors13, yet membrane transport applications of short CNTs remain underexplored. Here we show that short CNTs spontaneously insert into lipid bilayers and live cell membranes to form channels that exhibit a unitary conductance of 70–100 picosiemens under physiological conditions. Despite their structural simplicity, these ‘CNT porins’ transport water, protons, small ions and DNA, stochastically switch between metastable conductance substates, and display characteristic macromolecule-induced ionic current blockades. We also show that local channel and membrane charges can control the conductance and ion selectivity of the CNT porins, thereby establishing these nanopores as a promising biomimetic platform for developing cell interfaces, studying transport in biological channels, and creating stochastic sensors.

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References

  1. 1.

    , , , & Structural basis of water-specific transport through the AQP1 water channel. Nature 414, 872–878 (2001)

  2. 2.

    et al. Synthetic lipid membrane channels formed by designed DNA nanostructures. Science 338, 932–936 (2012)

  3. 3.

    et al. Ion-beam sculpting at nanometre length scales. Nature 412, 166–169 (2001)

  4. 4.

    et al. Aligned multiwalled carbon nanotube membranes. Science 303, 62–65 (2004)

  5. 5.

    et al. Translocation of single-stranded DNA through single-walled carbon nanotubes. Science 327, 64–67 (2010)

  6. 6.

    , , & Coherence resonance in a single-walled carbon nanotube ion channel. Science 329, 1320–1324 (2010)

  7. 7.

    & Simulations of electrophoretic RNA transport through transmembrane carbon nanotubes. Biophys. J. 94, 2546–2557 (2008)

  8. 8.

    , & Water conduction through the hydrophobic channel of a carbon nanotube. Nature 414, 188–190 (2001)

  9. 9.

    , , & Understanding nature's design for a nanosyringe. Proc. Natl Acad. Sci. USA 101, 4431–4434 (2004)

  10. 10.

    , , , & Interactions of end-functionalized nanotubes with lipid vesicles: spontaneous insertion and nanotube self-organization. Curr. Nanosci. 7, 699–715 (2011)

  11. 11.

    et al. How do functionalized carbon nanotubes land on, bind to and pierce through model and plasma membranes. Nanoscale 5, 10242–10250 (2013)

  12. 12.

    , , & Carbon nanotubes as multifunctional biological transporters and near-infrared agents for selective cancer cell destruction. Proc. Natl Acad. Sci. USA 102, 11600–11605 (2005)

  13. 13.

    , , , & Ultrashort single-walled carbon nanotubes in a lipid bilayer as a new nanopore sensor. Nature Commun. 4, (2013)

  14. 14.

    et al. Optical properties of ultrashort semiconducting single-walled carbon nanotube capsules down to sub-10 nm. J. Am. Chem. Soc. 130, 6551–6555 (2008)

  15. 15.

    et al. Imidazole-quartet water and proton dipolar channels. Angew. Chem. Int. Ed. 50, 11366–11372 (2011)

  16. 16.

    et al. Ion exclusion by sub 2-nm carbon nanotube pores. Proc. Natl Acad. Sci. USA 105, 17250–17255 (2008)

  17. 17.

    , , , & Barriers to superfast water transport in carbon nanotube membranes. Nano Lett. 13, 1910–1914 (2013)

  18. 18.

    & Interaction of the noncovalent molecular adapter, β-cyclodextrin, with the staphylococcal α-hemolysin pore. Biophys. J. 79, 1967–1975 (2000)

  19. 19.

    , & Residue ionization and ion transport through OmpF channels. Biophys. J. 85, 3718–3729 (2003)

  20. 20.

    et al. Nanoprecipitation-assisted ion current oscillations. Nature Nanotechnol. 3, 51–57 (2007)

  21. 21.

    , , , & Electric-field-induced wetting and dewetting in single hydrophobic nanopores. Nature Nanotechnol. 6, 798–802 (2011)

  22. 22.

    et al. Rapid switching of ion current in narrow pores: implications for biological ion channels. Proc. R. Soc. Lond. B 252, 187–192 (1993)

  23. 23.

    et al. Stochastic pore blocking and gating in PDMS–glass nanopores from vapor–liquid phase transitions. J. Phys. Chem. C 117, 9641–9651 (2013)

  24. 24.

    , & Ionic capillary evaporation in weakly charged nanopores. Phys. Rev. Lett. 105, 158103 (2010)

  25. 25.

    , , & Characterization of individual polynucleotide molecules using a membrane channel. Proc. Natl Acad. Sci. USA 93, 13770–13773 (1996)

  26. 26.

    et al. Hybrid pore formation by directed insertion of α-haemolysin into solid-state nanopores. Nature Nanotechnol. 5, 874–877 (2010)

  27. 27.

    & Nanopore sensors for nucleic acid analysis. Nature Nanotechnol. 6, 615–624 (2011)

  28. 28.

    , & Effect of tip functionalization on transport through vertically oriented carbon nanotube membranes. J. Am. Chem. Soc. 127, 9062–9070 (2005)

  29. 29.

    , , & Incorporation of a viral DNA-packaging motor channel in lipid bilayers for real-time, single-molecule sensing of chemicals and double-stranded DNA. Nature Protocols 8, 373–392 (2013)

  30. 30.

    , & Ionic selectivity, saturation, and block in gramicidin A channels. J. Membr. Biol. 40, 97–116 (1978)

  31. 31.

    , , , & Shape bistability of a membrane neck: a toggle switch to control vesicle content release. Proc. Natl Acad. Sci. USA 100, 8698–8703 (2003)

  32. 32.

    et al. Geometric catalysis of membrane fission driven by flexible dynamin rings. Science 339, 1433–1436 (2013)

  33. 33.

    , , , & Computer control of microscopes using µManager. Curr. Protoc. Molec. Biol. 92, 14.20.1–14.20.17 (2010)

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Acknowledgements

Parts of this work were supported by the US Department of Energy, Office of Basic Energy Sciences, Division of Materials Sciences and Engineering (characterization and transport studies), and the LDRD programme at LLNL, 12-ERD-073 (synthesis). R.T. acknowledges support from the LSP programme at LLNL. D.M. acknowledges support from an ROTC summer fellowship. V.A.F. acknowledges partial support by the Spanish Ministry of Economy and Competitiveness, grant BFU2012-34885, co-financed with European FEDER funds, and the Basque Government, grant IE12-332. Work at LLNL was performed under the auspices of the US Department of Energy under contract DE-AC52-07NA27344. Work at the Molecular Foundry was supported by the Office of Science, Office of Basic Energy Sciences, of the US Department of Energy under contract DE-AC02-05CH11231.

Author information

Author notes

    • Jia Geng
    •  & Kyunghoon Kim

    These authors contributed equally to this work.

Affiliations

  1. Biology and Biotechnology Division, Physical and Life Sciences Directorate, Lawrence Livermore National Laboratory, Livermore, California 94550, USA

    • Jia Geng
    • , Kyunghoon Kim
    • , Ramya Tunuguntla
    • , Kang Rae Cho
    • , Dayannara Munoz
    •  & Aleksandr Noy
  2. School of Natural Sciences, University of California, Merced, California 95340, USA

    • Jia Geng
    • , Jianfei Zhang
    •  & Aleksandr Noy
  3. The Molecular Foundry, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA

    • Jia Geng
    • , Kyunghoon Kim
    • , Ramya Tunuguntla
    • , Caroline M. Ajo-Franklin
    •  & Aleksandr Noy
  4. Mechanical Engineering Department, University of California, Berkeley, California 94720, USA

    • Kyunghoon Kim
    •  & Costas P. Grigoropoulos
  5. Biophysics Unit (CSIC, UPV/EHU) and Department of Biochemistry and Molecular Biology, University of the Basque Country, 48940 Leioa, Spain

    • Artur Escalada
    • , Anna V. Shnyrova
    •  & Vadim A. Frolov
  6. Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA

    • Luis R. Comolli
  7. Department of Materials Science and Engineering, University of California, Berkeley, California 94720, USA

    • Frances I. Allen
  8. Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA

    • Frances I. Allen
    •  & Caroline M. Ajo-Franklin
  9. National Center for Electron Microscopy, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA

    • Frances I. Allen
  10. Materials Science Division, Lawrence Livermore National Laboratory, Livermore, California 94550, USA

    • Y. Morris Wang
  11. Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA

    • Caroline M. Ajo-Franklin
  12. Ikerbasque, Basque Foundation for Science, 48011 Bilbao, Spain

    • Vadim A. Frolov

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Contributions

J.G. performed and analysed (with D.M.) the planar lipid bilayer transport measurements; K.K. performed bulk transport studies; J.Z., K.K., J.G. and R.T. performed the synthesis and purification, K.R.C. performed AFM analysis, Y.M.W., L.R.C., F.I.A. and K.K. performed TEM analysis; L.R.C. and F.I.A. performed cryo-TEM analysis; A.N. conceived and directed the research, and wrote the manuscript draft; A.E., A.V.S. and V.A.F. designed the cell and GUV reconstitution experiments, and A.E. and A.V.S. performed them. All authors contributed to the data analysis, discussion, and manuscript preparation.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Aleksandr Noy.

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DOI

https://doi.org/10.1038/nature13817

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