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Bilayer Reconstitution of Voltage-Dependent Ion Channels using a Microfabricated Silicon Chip

Pantoja, Rigo and Sigg, Daniel and Blunck, Rikard and Bezanilla, Francisco and Heath, James R. (2001) Bilayer Reconstitution of Voltage-Dependent Ion Channels using a Microfabricated Silicon Chip. Biophysical Journal, 81 (4). pp. 2389-2394. ISSN 0006-3495. PMCID PMC1301709. https://resolver.caltech.edu/CaltechAUTHORS:20190513-153906998

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Abstract

Painted bilayers containing reconstituted ion channels serve as a well defined model system for electrophysiological investigations of channel structure and function. Horizontally oriented bilayers with easy solution access to both sides were obtained by painting a phospholipid:decane mixture across a cylindrical pore etched into a 200-μm thick silicon wafer. Silanization of the SiO_2 layer produced a hydrophobic surface that promoted the adhesion of the lipid mixture. Standard lithographic techniques and anisotropic deep-reactive ion etching were used to create pores with diameters from 50 to 200 μm. The cylindrical structure of the pore in the partition and the surface treatment resulted in stable bilayers. These were used to reconstitute Maxi K channels in the 100- and 200-μm diameter pores. The electrophysiological characteristics of bilayers suspended in microchips were comparable with that of other bilayer preparations. The horizontal orientation and good voltage clamping properties make the microchip bilayer method an excellent system to study the electrical properties of reconstituted membrane proteinssimultaneously with optical probes.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/s0006-3495(01)75885-7DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1301709PubMed CentralArticle
ORCID:
AuthorORCID
Blunck, Rikard0000-0003-4484-2907
Heath, James R.0000-0001-5356-4385
Additional Information:© 2001 The Biophysical Society. Published by Elsevier Inc. Received 13 February 2001, Revised 12 July 2001, Available online 6 January 2009. We thank Dr. Riccardo Olcese for invaluable discussions with regard to single-channel analysis and Dr. Ligia Toro for providing us with the Maxi K clone. We also thank Dr. Ramon Latorre for very useful bilayer discussions. We also gratefully acknowledge the assistance provided by the UCLA Nanoelectronics Research Facility staff. This research was supported by the William M. Keck Foundation and National Institutes of Health Grant GM 30376.
Funders:
Funding AgencyGrant Number
W. M. Keck FoundationUNSPECIFIED
NIHGM30376
Issue or Number:4
PubMed Central ID:PMC1301709
Record Number:CaltechAUTHORS:20190513-153906998
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20190513-153906998
Official Citation:Rigo Pantoja, Daniel Sigg, Rikard Blunck, Francisco Bezanilla, James R. Heath, Bilayer Reconstitution of Voltage-Dependent Ion Channels using a Microfabricated Silicon Chip, Biophysical Journal, Volume 81, Issue 4, 2001, Pages 2389-2394, ISSN 0006-3495, https://doi.org/10.1016/S0006-3495(01)75885-7. (http://www.sciencedirect.com/science/article/pii/S0006349501758857)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:95454
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:14 May 2019 00:21
Last Modified:03 Oct 2019 21:13

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