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Unlocking the Reversal Potential of Solid Supported Membrane Electrophysiology to Determine Transport Stoichiometry

View ORCID ProfileNathan E. Thomas, View ORCID ProfileKatherine A. Henzler-Wildman
doi: https://doi.org/10.1101/2020.05.07.082438
Nathan E. Thomas
Department of Biochemistry, University of Wisconsin-Madison
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Katherine A. Henzler-Wildman
Department of Biochemistry, University of Wisconsin-Madison
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  • For correspondence: henzlerwildm@wisc.edu
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Abstract

Transport stoichiometry provides insight into the mechanism and function of ion-coupled transporters, but measuring transport stoichiometry is time-consuming and technically difficult. With the increasing evidence that many ion-coupled transporters employ multiple transport stoichiometries under different conditions, improved methods to determine transport stoichiometry are required to accurately characterize transporter activity. Reversal potential was previously shown to be a reliable, general method for determining the transport stoichiometry of ion-coupled transporters (Fitzgerald & Mindell, 2017). Here, we develop a new technique for measuring transport stoichiometry with greatly improved throughput using solid supported membrane electrophysiology (SSME). Using this technique, we are able to verify the recent report of a fixed 2:1 stoichiometry for the proton:guanidinium antiporter Gdx. Our SSME method requires only small amounts of transporter and provides a fast, easy, general method for measuring transport stoichiometry, which will facilitate future mechanistic and functional studies of ion-coupled transporters.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted May 07, 2020.
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Unlocking the Reversal Potential of Solid Supported Membrane Electrophysiology to Determine Transport Stoichiometry
Nathan E. Thomas, Katherine A. Henzler-Wildman
bioRxiv 2020.05.07.082438; doi: https://doi.org/10.1101/2020.05.07.082438
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Unlocking the Reversal Potential of Solid Supported Membrane Electrophysiology to Determine Transport Stoichiometry
Nathan E. Thomas, Katherine A. Henzler-Wildman
bioRxiv 2020.05.07.082438; doi: https://doi.org/10.1101/2020.05.07.082438

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