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

Hartel, Andreas J. W. ; Ong, Peijie ; Schroeder, Indra ; Giese, M. Hunter ; Shekar, Siddharth ; Clarke, Oliver B. ; Zalk, Ran ; Marks, Andrew R. ; Hendrickson, Wayne A. ; Shepard, Kenneth L. (2018)
Single-channel recordings of RyR1 at microsecond resolution in CMOS-suspended membranes.
In: Proceedings of the National Academy of Sciences, 115 (8)
doi: 10.1073/pnas.1712313115
Article, Bibliographie

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. ; Ong, Peijie ; Schroeder, Indra ; Giese, M. Hunter ; Shekar, Siddharth ; Clarke, Oliver B. ; Zalk, Ran ; Marks, Andrew R. ; Hendrickson, Wayne A. ; Shepard, Kenneth L.
Type of entry: Bibliographie
Title: Single-channel recordings of RyR1 at microsecond resolution in CMOS-suspended membranes
Language: English
Date: 2018
Publisher: National Academy of Sciences
Journal or Publication Title: Proceedings of the National Academy of Sciences
Volume of the journal: 115
Issue Number: 8
DOI: 10.1073/pnas.1712313115
URL / URN: https://doi.org/10.1073/pnas.1712313115
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.

Divisions: 10 Department of Biology > Plant Membrane Biophyscis (20.12.23 renamed in Biology of Algae and Protozoa)
10 Department of Biology
Date Deposited: 23 Feb 2018 07:42
Last Modified: 23 Feb 2018 07:42
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