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Single-channel recordings of RyR1 at microsecond resolution in CMOS-suspended membranes

Hartel, Andreas J. W. and Ong, Peijie and Schroeder, Indra and Giese, M. Hunter and Shekar, Siddharth and Clarke, Oliver B. and Zalk, Ran and Marks, Andrew R. and Hendrickson, Wayne A. and Shepard, Kenneth L. (2018):
Single-channel recordings of RyR1 at microsecond resolution in CMOS-suspended membranes.
115, In: Proceedings of the National Academy of Sciences, (8), National Academy of Sciences, pp. E1789-E1798, ISSN 0027-8424, DOI: 10.1073/pnas.1712313115,
[Online-Edition: https://doi.org/10.1073/pnas.1712313115],
[Article]

Abstract

Single-channel recordings are widely used to explore functional properties of ion channels. Typically, such recordings are performed at bandwidths of less than 10 kHz because of signal-to-noise considerations, limiting the temporal resolution available for studying fast gating dynamics to greater than 100 µs. Here we present experimental methods that directly integrate suspended lipid bilayers with high-bandwidth, low-noise transimpedance amplifiers based on complementary metal-oxide-semiconductor (CMOS) integrated circuits (IC) technology to achieve bandwidths in excess of 500 kHz and microsecond temporal resolution. We use this CMOS-integrated bilayer system to study the type 1 ryanodine receptor (RyR1), a Ca2+-activated intracellular Ca2+-release channel located on the sarcoplasmic reticulum. We are able to distinguish multiple closed states not evident with lower bandwidth recordings, suggesting the presence of an additional Ca2+ binding site, distinct from the site responsible for activation. An extended beta distribution analysis of our high-bandwidth data can be used to infer closed state flicker events as fast as 35 ns. These events are in the range of single-file ion translocations.

Item Type: Article
Erschienen: 2018
Creators: Hartel, Andreas J. W. and Ong, Peijie and Schroeder, Indra and Giese, M. Hunter and Shekar, Siddharth and Clarke, Oliver B. and Zalk, Ran and Marks, Andrew R. and Hendrickson, Wayne A. and Shepard, Kenneth L.
Title: Single-channel recordings of RyR1 at microsecond resolution in CMOS-suspended membranes
Language: English
Abstract:

Single-channel recordings are widely used to explore functional properties of ion channels. Typically, such recordings are performed at bandwidths of less than 10 kHz because of signal-to-noise considerations, limiting the temporal resolution available for studying fast gating dynamics to greater than 100 µs. Here we present experimental methods that directly integrate suspended lipid bilayers with high-bandwidth, low-noise transimpedance amplifiers based on complementary metal-oxide-semiconductor (CMOS) integrated circuits (IC) technology to achieve bandwidths in excess of 500 kHz and microsecond temporal resolution. We use this CMOS-integrated bilayer system to study the type 1 ryanodine receptor (RyR1), a Ca2+-activated intracellular Ca2+-release channel located on the sarcoplasmic reticulum. We are able to distinguish multiple closed states not evident with lower bandwidth recordings, suggesting the presence of an additional Ca2+ binding site, distinct from the site responsible for activation. An extended beta distribution analysis of our high-bandwidth data can be used to infer closed state flicker events as fast as 35 ns. These events are in the range of single-file ion translocations.

Journal or Publication Title: Proceedings of the National Academy of Sciences
Volume: 115
Number: 8
Publisher: National Academy of Sciences
Divisions: 10 Department of Biology > Plant Membrane Biophysics
10 Department of Biology
Date Deposited: 23 Feb 2018 07:42
DOI: 10.1073/pnas.1712313115
Official URL: https://doi.org/10.1073/pnas.1712313115
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