Probing Ion Configurations in the KcsA Selectivity Filter with Single-Isotope Labels and 2D IR Spectroscopy

Author(s)Ryan, Matthew J.
Author(s)Gao, Lujia
Author(s)Valiyaveetil, Francis I.
Author(s)Zanni, Martin T.
Author(s)Kananenka, Alexei A.
Date Accessioned2023-11-10T16:46:32Z
Date Available2023-11-10T16:46:32Z
Publication Date2023-08-23
DescriptionThis article was originally published in Journal of the American Chemical Society. The version of record is available at: https://doi.org/10.1021/jacs.3c05339
AbstractThe potassium ion (K+) configurations of the selectivity filter of the KcsA ion channel protein are investigated with two-dimensional infrared (2D IR) spectroscopy of amide I vibrations. Single 13C–18O isotope labels are used, for the first time, to selectively probe the S1/S2 or S2/S3 binding sites in the selectivity filter. These binding sites have the largest differences in ion occupancy in two competing K+ transport mechanisms: soft-knock and hard-knock. According to the former, water molecules alternate between K+ ions in the selectivity filter while the latter assumes that K+ ions occupy the adjacent sites. Molecular dynamics simulations and computational spectroscopy are employed to interpret experimental 2D IR spectra. We find that in the closed conductive state of the KcsA channel, K+ ions do not occupy adjacent binding sites. The experimental data is consistent with simulated 2D IR spectra of soft-knock ion configurations. In contrast, the simulated spectra for the hard-knock ion configurations do not reproduce the experimental results. 2D IR spectra of the hard-knock mechanism have lower frequencies, homogeneous 2D lineshapes, and multiple peaks. In contrast, ion configurations of the soft-knock model produce 2D IR spectra with a single peak at a higher frequency and inhomogeneous lineshape. We conclude that under equilibrium conditions, in the absence of transmembrane voltage, both water and K+ ions occupy the selectivity filter of the KcsA channel in the closed conductive state. The ion configuration is central to the mechanism of ion transport through potassium channels.
SponsorA.A.K. acknowledges the support from NSF via award number OIA-2229651 and start-up funds provided by the College of Arts and Sciences and the Department of Physics and Astronomy of the University of Delaware. M.T.Z. and F.I.V. acknowledge the financial support from the National Institutes of Health grant R01GM135936. Calculations were performed with high-performance computing resources provided by the University of Delaware.
CitationRyan, Matthew J., Lujia Gao, Francis I. Valiyaveetil, Martin T. Zanni, and Alexei A. Kananenka. “Probing Ion Configurations in the KcsA Selectivity Filter with Single-Isotope Labels and 2D IR Spectroscopy.” Journal of the American Chemical Society 145, no. 33 (August 23, 2023): 18529–37. https://doi.org/10.1021/jacs.3c05339.
ISSN1520-5126
URLhttps://udspace.udel.edu/handle/19716/33625
Languageen_US
PublisherJournal of the American Chemical Society
dc.rightshttps://creativecommons.org/licenses/by-nc-nd/4.0/
Keywordsinfrared light
Keywordsions
Keywordspeptides and proteins
Keywordspotassium
Keywordsselectivity
TitleProbing Ion Configurations in the KcsA Selectivity Filter with Single-Isotope Labels and 2D IR Spectroscopy
TypeArticle
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