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

Tunuguntla, Ramya H.; Escalada, Artur; A Frolov, Vadim; Noy, Aleksandr

doi:10.1038/nprot.2016.119

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

Journal Article · Thu Sep 22 00:00:00 EDT 2016 · Nature Protocols

DOI:https://doi.org/10.1038/nprot.2016.119· OSTI ID:1530684

Tunuguntla, Ramya H. ^[1]; Escalada, Artur ^[2]; A Frolov, Vadim ^[3]; Noy, Aleksandr ^[4]

Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Univ. of Basque Country, Leioa (Spain)
Univ. of Basque Country, Leioa (Spain); Basque Foundation for Science, Bilbao (Spain)
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Univ. of California Merced, Merced, CA (United States)

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 report on 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. Moreover, 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.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)

Sponsoring Organization:: Spanish Ministry of Economy and Competitiveness; USDOE National Nuclear Security Administration (NNSA); USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22). Materials Sciences & Engineering Division

Grant/Contract Number:: AC02-05CH11231; AC52-07NA27344

OSTI ID:: 1530684

Report Number(s):: LLNL-JRNL--691260; 819066

Journal Information:: Nature Protocols, Journal Name: Nature Protocols Journal Issue: 10 Vol. 11; ISSN 1754-2189

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

References (45)

Bioelectronic Light-Gated Transistors with Biologically Tunable Performance Tunuguntla, Ramya H.; Bangar, Mangesh A.; Kim, Kyunghoon Advanced Materials, Vol. 27, Issue 5 https://doi.org/10.1002/adma.201403988	journal	November 2014
Imidazole-Quartet Water and Proton Dipolar Channels Le Duc, Yann; Michau, Mathieu; Gilles, Arnaud Angewandte Chemie International Edition, Vol. 50, Issue 48 https://doi.org/10.1002/anie.201103312	journal	August 2011
Micro- and Nano-Technologies for Lipid Bilayer-Based Ion-Channel Functional Assays Hirano-Iwata, Ayumi; Ishinari, Yutaka; Yamamoto, Hideaki Chemistry - An Asian Journal, Vol. 10, Issue 6 https://doi.org/10.1002/asia.201403391	journal	February 2015
Single-Channel Recording Sakmann, Bert; Neher, Erwin https://doi.org/10.1007/978-1-4615-7858-1	book	January 1983
Perforated Whole-Cell Patch-Clamp Recording Linley, John E. Ion Channels. Methods in Molecular Biology https://doi.org/10.1007/978-1-62703-351-0_11	book	January 2013
Selective suspension in aqueous sodium dodecyl sulfate according to electronic structure type allows simple separation of metallic from semiconducting single-walled carbon nanotubes Moshammer, Kai; Hennrich, Frank; Kappes, Manfred M. Nano Research, Vol. 2, Issue 8 https://doi.org/10.1007/s12274-009-9057-0	journal	August 2009
Molecular Dynamics of Synthetic Leucine-Serine Ion Channels in a Phospholipid Membrane Randa, Holly S.; Forrest, Lucy R.; Voth, Gregory A. Biophysical Journal, Vol. 77, Issue 5 https://doi.org/10.1016/S0006-3495(99)77077-3	journal	November 1999
Nanofluidics in carbon nanotubes Noy, Aleksandr; Park, Hyung Gyu; Fornasiero, Francesco Nano Today, Vol. 2, Issue 6 https://doi.org/10.1016/S1748-0132(07)70170-6	journal	December 2007
Biomimetic membranes: A review Shen, Yue-xiao; Saboe, Patrick O.; Sines, Ian T. Journal of Membrane Science, Vol. 454 https://doi.org/10.1016/j.memsci.2013.12.019	journal	March 2014
Single Carbon Nanotube Membranes: A Well-Defined Model for Studying Mass Transport through Nanoporous Materials Sun, Li; Crooks, Richard M. Journal of the American Chemical Society, Vol. 122, Issue 49, p. 12340-12345 https://doi.org/10.1021/ja002429w	journal	December 2000
Optical Properties of Ultrashort Semiconducting Single-Walled Carbon Nanotube Capsules Down to Sub-10 nm Sun, Xiaoming; Zaric, Sasa; Daranciang, Dan Journal of the American Chemical Society, Vol. 130, Issue 20 https://doi.org/10.1021/ja8006929	journal	May 2008
Ion Transport in Nanofluidic Channels Daiguji, Hirofumi; Yang, Peidong; Majumdar, Arun Nano Letters, Vol. 4, Issue 1 https://doi.org/10.1021/nl0348185	journal	January 2004
Enrichment of Single-Walled Carbon Nanotubes by Diameter in Density Gradients Arnold, Michael S.; Stupp, Samuel I.; Hersam, Mark C. Nano Letters, Vol. 5, Issue 4 https://doi.org/10.1021/nl050133o	journal	April 2005
Separation of Single-Walled Carbon Nanotubes with a Gel Permeation Chromatography System Flavel, Benjamin S.; Moore, Katherine E.; Pfohl, Moritz ACS Nano, Vol. 8, Issue 2 https://doi.org/10.1021/nn4062116	journal	January 2014
Stochastic sensing of organic analytes by a pore-forming protein containing a molecular adapter Gu, Li-Qun; Braha, Orit; Conlan, Sean Nature, Vol. 398, Issue 6729 https://doi.org/10.1038/19491	journal	April 1999
Reconstitution of Cell Membrane Structure in vitro and its Transformation into an Excitable System Mueller, Paul; Rudin, Donald O.; Ti Tien, H. Nature, Vol. 194, Issue 4832 https://doi.org/10.1038/194979a0	journal	June 1962
The appearance of single-ion channels in unmodified lipid bilayer membranes at the phase transition temperature Antonov, V. F.; Petrov, V. V.; Molnar, A. A. Nature, Vol. 283, Issue 5747 https://doi.org/10.1038/283585a0	journal	February 1980
A voltage-dependent channel involved in nutrient uptake by red blood cells infected with the malaria parasite Desai, Sanjay A.; Bezrukov, Sergey M.; Zimmerberg, Joshua Nature, Vol. 406, Issue 6799 https://doi.org/10.1038/35023000	journal	August 2000
Water conduction through the hydrophobic channel of a carbon nanotube Hummer, G.; Rasaiah, J. C.; Noworyta, J. P. Nature, Vol. 414, Issue 6860 https://doi.org/10.1038/35102535	journal	November 2001
Enhanced flow in carbon nanotubes Majumder, Mainak; Chopra, Nitin; Andrews, Rodney Nature, Vol. 438, Issue 7064 https://doi.org/10.1038/438044a	journal	November 2005
Stochastic transport through carbon nanotubes in lipid bilayers and live cell membranes Geng, Jia; Kim, Kyunghoon; Zhang, Jianfei Nature, Vol. 514, Issue 7524 https://doi.org/10.1038/nature13817	journal	October 2014
A polysaccharide bioprotonic field-effect transistor Zhong, Chao; Deng, Yingxin; Roudsari, Anita Fadavi Nature Communications, Vol. 2, Issue 1 https://doi.org/10.1038/ncomms1489	journal	September 2011
Ultrashort single-walled carbon nanotubes in a lipid bilayer as a new nanopore sensor Liu, Lei; Yang, Chun; Zhao, Kai Nature Communications, Vol. 4, Issue 1 https://doi.org/10.1038/ncomms3989	journal	December 2013
Sorting carbon nanotubes by electronic structure using density differentiation Arnold, Michael S.; Green, Alexander A.; Hulvat, James F. Nature Nanotechnology, Vol. 1, Issue 1, p. 60-65 https://doi.org/10.1038/nnano.2006.52	journal	October 2006
Solid-state nanopores Dekker, Cees Nature Nanotechnology, Vol. 2, Issue 4 https://doi.org/10.1038/nnano.2007.27	journal	March 2007
Ultrafast proton transport in sub-1-nm diameter carbon nanotube porins Tunuguntla, Ramya H.; Allen, Frances I.; Kim, Kyunghoon Nature Nanotechnology, Vol. 11, Issue 7 https://doi.org/10.1038/nnano.2016.43	journal	April 2016
Single-channel recording of ligand-gated ion channels Mortensen, Martin; Smart, Trevor G. Nature Protocols, Vol. 2, Issue 11 https://doi.org/10.1038/nprot.2007.403	journal	November 2007
Protein reconstitution into freestanding planar lipid membranes for electrophysiological characterization Gutsmann, Thomas; Heimburg, Thomas; Keyser, Ulrich Nature Protocols, Vol. 10, Issue 1 https://doi.org/10.1038/nprot.2015.003	journal	December 2014
Synthetic ion channels in bilayer membranes Fyles, Thomas M. Chem. Soc. Rev., Vol. 36, Issue 2 https://doi.org/10.1039/B603256G	journal	January 2007
Miniaturised technologies for the development of artificial lipid bilayer systems Zagnoni, Michele Lab on a Chip, Vol. 12, Issue 6 https://doi.org/10.1039/c2lc20991h	journal	January 2012
Bioelectronic silicon nanowire devices using functional membrane proteins Misra, N.; Martinez, J. A.; Huang, S. -C. J. Proceedings of the National Academy of Sciences, Vol. 106, Issue 33 https://doi.org/10.1073/pnas.0904850106	journal	August 2009
Shape bistability of a membrane neck: A toggle switch to control vesicle content release Frolov, V. A.; Lizunov, V. A.; Dunina-Barkovskaya, A. Y. Proceedings of the National Academy of Sciences, Vol. 100, Issue 15 https://doi.org/10.1073/pnas.1432962100	journal	July 2003
The Aquaporins, Blueprints for Cellular Plumbing Systems Agre, Peter; Bonhivers, Mélanie; Borgnia, Mario J. Journal of Biological Chemistry, Vol. 273, Issue 24 https://doi.org/10.1074/jbc.273.24.14659	journal	June 1998
Water, proton, and ion transport: from nanotubes to proteins Hummer, Gerhard Molecular Physics, Vol. 105, Issue 2-3 https://doi.org/10.1080/00268970601140784	journal	January 2007
Carbon nanotubes Dresselhaus, Mildred; Dresselhaus, Gene; Eklund, Peter Physics World, Vol. 11, Issue 1 https://doi.org/10.1088/2058-7058/11/1/32	journal	January 1998
Proton Transport through Water-Filled Carbon Nanotubes Dellago, Christoph; Naor, Mor M.; Hummer, Gerhard Physical Review Letters, Vol. 90, Issue 10 https://doi.org/10.1103/PhysRevLett.90.105902	journal	March 2003
Kinetics and Mechanism of Proton Transport across Membrane Nanopores Dellago, Christoph; Hummer, Gerhard Physical Review Letters, Vol. 97, Issue 24 https://doi.org/10.1103/PhysRevLett.97.245901	journal	December 2006
Aligned Multiwalled Carbon Nanotube Membranes Hinds, B. J. Science, Vol. 303, Issue 5654 https://doi.org/10.1126/science.1092048	journal	January 2004
Fast Mass Transport Through Sub-2-Nanometer Carbon Nanotubes Holt, J. K. Science, Vol. 312, Issue 5776 https://doi.org/10.1126/science.1126298	journal	May 2006
Translocation of Single-Stranded DNA Through Single-Walled Carbon Nanotubes Liu, H.; He, J.; Tang, J. Science, Vol. 327, Issue 5961 https://doi.org/10.1126/science.1181799	journal	December 2009
Coherence Resonance in a Single-Walled Carbon Nanotube Ion Channel Lee, Chang Young; Choi, Wonjoon; Han, Jae-Hee Science, Vol. 329, Issue 5997 https://doi.org/10.1126/science.1193383	journal	September 2010
The Structure of the Potassium Channel: Molecular Basis of K+ Conduction and Selectivity Doyle, D. A. Science, Vol. 280, Issue 5360 https://doi.org/10.1126/science.280.5360.69	journal	April 1998
Proton Conductivity of Nafion 117 as Measured by a Four-Electrode AC Impedance Method Sone, Yoshitsugu Journal of The Electrochemical Society, Vol. 143, Issue 4 https://doi.org/10.1149/1.1836625	journal	January 1996
Aquaporin water channels: atomic structure molecular dynamics meet clinical medicine Kozono, David; Yasui, Masato; King, Landon S. Journal of Clinical Investigation, Vol. 109, Issue 11 https://doi.org/10.1172/JCI0215851	journal	June 2002
Architecture and Selectivity in Aquaporins: 2.5 Å X-Ray Structure of Aquaporin Z Savage, David F.; Egea, Pascal F.; Robles-Colmenares, Yaneth PLoS Biology, Vol. 1, Issue 3 https://doi.org/10.1371/journal.pbio.0000072	journal	December 2003

Cited By (4)

Carbon Nanotube Porins in Amphiphilic Block Copolymers as Fully Synthetic Mimics of Biological Membranes Sanborn, Jeremy R.; Chen, Xi; Yao, Yun-Chiao Advanced Materials, Vol. 30, Issue 51 https://doi.org/10.1002/adma.201803355	journal	October 2018
Impact of PEG additives and pore rim functionalization on water transport through sub-1 nm carbon nanotube porins Tunuguntla, Ramya H.; Hu, Andrew Y.; Zhang, Yuliang Faraday Discussions, Vol. 209 https://doi.org/10.1039/c8fd00068a	journal	January 2018
Versatile cyclodextrin nanotube synthesis with functional anchors for efficient ion channel formation: design, characterization and ion conductance Mamad-Hemouch, Hajar; Bacri, Laurent; Huin, Cécile Nanoscale, Vol. 10, Issue 32 https://doi.org/10.1039/c8nr02623h	journal	January 2018
Enhanced water permeability and tunable ion selectivity in subnanometer carbon nanotube porins Tunuguntla, Ramya H.; Henley, Robert Y.; Yao, Yun-Chiao Science, Vol. 357, Issue 6353 https://doi.org/10.1126/science.aan2438	journal	August 2017

Similar Records

Structure of Carbon Nanotube Porins in Lipid Bilayers: An in Situ Small-Angle X-ray Scattering (SAXS) Study [Atomic-level structure of carbon nanotube porins in lipid bilayers: an in-situ small-angle x-ray scattering (SAXS) study]

Journal Article · Sun Jun 19 20:00:00 EDT 2016 · Nano Letters · OSTI ID:1367965

Sonochemical Synthesis and Ion Transport Properties of Surfactant-Stabilized Carbon Nanotube Porins

Journal Article · Wed Oct 11 20:00:00 EDT 2023 · Journal of Physical Chemistry Letters · OSTI ID:2229991

Carbon nanotube porin diffusion in mixed composition supported lipid bilayers

Journal Article · Thu Jul 16 20:00:00 EDT 2020 · Scientific Reports · OSTI ID:1638906

Related Subjects

59 BASIC BIOLOGICAL SCIENCES

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

Citation Formats

References (45)

Cited By (4)

Similar Records

Related Subjects