Chloride binding and cholesterol effects on high frequency complex nonlinear capacitance (cNLC) in the mouse outer hair cell: experiment and molecular dynamics. Bai, J., Zhang, C., Renigunta, V., Oliver, D., Navaratnam, D. S., Beckstein, O., & Santos-Sacchi, J. January, 2024. Pages: 2024.01.29.577264 Section: New Results![Chloride binding and cholesterol effects on high frequency complex nonlinear capacitance (cNLC) in the mouse outer hair cell: experiment and molecular dynamics [link]](https://bibbase.org/img/filetypes/link.svg "link")
Paper doi abstract bibtex The function of prestin (SLC26a5), an anion transport family member, has evolved to enhance auditory sensitivity and frequency selectivity by providing mechanical feedback via outer hair cells (OHC) into the organ of Corti. The frequency extent of this boost is governed by the voltage-dependent kinetics of the protein charge movements, otherwise known as nonlinear capacitance (NLC) that we measure in membrane patches under voltage clamp. Here we extend our previous studies on guinea pig OHCs by studying the frequency response of NLC in the mouse OHC, a species with higher frequency auditory needs. We find that the characteristic frequency cut-off (Fis) for the mouse surpasses that of the guinea pig, being 27 kHz vs. 19 kHz, respectively; nevertheless, each shows significant activity in the ultrasonic range. We also evaluate the influence of anion binding on prestin frequency response. Several single point mutations within the chloride binding pocket of prestin (e.g., S396E, S398E) lack anion influence. In agreement, we show absence of anion binding through molecular dynamics (MD) simulations. NLC Fis in the S396E knock-in mouse remains the same as controls, indicating that high frequency activity is likely governed by viscoelastic loads within the membrane characterized by stretched-exponential frequency roll-off. Accordingly, treatment with MBCD, which removes membrane cholesterol, possibly from prestin itself, and can alter membrane fluidity, augments NLC Fis out to 39 kHz. Although interactions between membrane lipid and prestin have been suggested from structural studies to arise at their interfacial boundaries within the membrane, our MD simulations suggest that phospholipids can insert within transmembrane domains of prestin during voltage perturbation. Such novel lipid-protein interactions could account for our observed changes in the phase of prestin voltage-sensor charge movements across frequency. We hypothesize that because prestin tertiary structures of all species studied to-date are indistinguishable, it is likely that any special auditory requirements of individual species for cochlear amplification have evolved to capitalize on prestin performance by modifying, not the protein itself, but the external loads on the protein, including those within the membrane and organ of Corti.
@misc{bai_chloride_2024,
title = {Chloride binding and cholesterol effects on high frequency complex nonlinear capacitance ({cNLC}) in the mouse outer hair cell: experiment and molecular dynamics},
copyright = {© 2024, Posted by Cold Spring Harbor Laboratory. The copyright holder for this pre-print is the author. All rights reserved. The material may not be redistributed, re-used or adapted without the author's permission.},
shorttitle = {Chloride binding and cholesterol effects on high frequency complex nonlinear capacitance ({cNLC}) in the mouse outer hair cell},
url = {https://www.biorxiv.org/content/10.1101/2024.01.29.577264v1},
doi = {10.1101/2024.01.29.577264},
abstract = {The function of prestin (SLC26a5), an anion transport family member, has evolved to enhance auditory sensitivity and frequency selectivity by providing mechanical feedback via outer hair cells (OHC) into the organ of Corti. The frequency extent of this boost is governed by the voltage-dependent kinetics of the protein charge movements, otherwise known as nonlinear capacitance (NLC) that we measure in membrane patches under voltage clamp. Here we extend our previous studies on guinea pig OHCs by studying the frequency response of NLC in the mouse OHC, a species with higher frequency auditory needs. We find that the characteristic frequency cut-off (Fis) for the mouse surpasses that of the guinea pig, being 27 kHz vs. 19 kHz, respectively; nevertheless, each shows significant activity in the ultrasonic range. We also evaluate the influence of anion binding on prestin frequency response. Several single point mutations within the chloride binding pocket of prestin (e.g., S396E, S398E) lack anion influence. In agreement, we show absence of anion binding through molecular dynamics (MD) simulations. NLC Fis in the S396E knock-in mouse remains the same as controls, indicating that high frequency activity is likely governed by viscoelastic loads within the membrane characterized by stretched-exponential frequency roll-off. Accordingly, treatment with MBCD, which removes membrane cholesterol, possibly from prestin itself, and can alter membrane fluidity, augments NLC Fis out to 39 kHz. Although interactions between membrane lipid and prestin have been suggested from structural studies to arise at their interfacial boundaries within the membrane, our MD simulations suggest that phospholipids can insert within transmembrane domains of prestin during voltage perturbation. Such novel lipid-protein interactions could account for our observed changes in the phase of prestin voltage-sensor charge movements across frequency. We hypothesize that because prestin tertiary structures of all species studied to-date are indistinguishable, it is likely that any special auditory requirements of individual species for cochlear amplification have evolved to capitalize on prestin performance by modifying, not the protein itself, but the external loads on the protein, including those within the membrane and organ of Corti.},
language = {en},
urldate = {2024-01-31},
publisher = {bioRxiv},
author = {Bai, Jun-Ping and Zhang, Chenou and Renigunta, Vijay and Oliver, Dominik and Navaratnam, Dhasakumar S. and Beckstein, Oliver and Santos-Sacchi, Joseph},
month = jan,
year = {2024},
note = {Pages: 2024.01.29.577264
Section: New Results},
}
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The material may not be redistributed, re-used or adapted without the author's permission.","shorttitle":"Chloride binding and cholesterol effects on high frequency complex nonlinear capacitance (cNLC) in the mouse outer hair cell","url":"https://www.biorxiv.org/content/10.1101/2024.01.29.577264v1","doi":"10.1101/2024.01.29.577264","abstract":"The function of prestin (SLC26a5), an anion transport family member, has evolved to enhance auditory sensitivity and frequency selectivity by providing mechanical feedback via outer hair cells (OHC) into the organ of Corti. The frequency extent of this boost is governed by the voltage-dependent kinetics of the protein charge movements, otherwise known as nonlinear capacitance (NLC) that we measure in membrane patches under voltage clamp. Here we extend our previous studies on guinea pig OHCs by studying the frequency response of NLC in the mouse OHC, a species with higher frequency auditory needs. We find that the characteristic frequency cut-off (Fis) for the mouse surpasses that of the guinea pig, being 27 kHz vs. 19 kHz, respectively; nevertheless, each shows significant activity in the ultrasonic range. We also evaluate the influence of anion binding on prestin frequency response. Several single point mutations within the chloride binding pocket of prestin (e.g., S396E, S398E) lack anion influence. In agreement, we show absence of anion binding through molecular dynamics (MD) simulations. NLC Fis in the S396E knock-in mouse remains the same as controls, indicating that high frequency activity is likely governed by viscoelastic loads within the membrane characterized by stretched-exponential frequency roll-off. Accordingly, treatment with MBCD, which removes membrane cholesterol, possibly from prestin itself, and can alter membrane fluidity, augments NLC Fis out to 39 kHz. Although interactions between membrane lipid and prestin have been suggested from structural studies to arise at their interfacial boundaries within the membrane, our MD simulations suggest that phospholipids can insert within transmembrane domains of prestin during voltage perturbation. Such novel lipid-protein interactions could account for our observed changes in the phase of prestin voltage-sensor charge movements across frequency. We hypothesize that because prestin tertiary structures of all species studied to-date are indistinguishable, it is likely that any special auditory requirements of individual species for cochlear amplification have evolved to capitalize on prestin performance by modifying, not the protein itself, but the external loads on the protein, including those within the membrane and organ of Corti.","language":"en","urldate":"2024-01-31","publisher":"bioRxiv","author":[{"propositions":[],"lastnames":["Bai"],"firstnames":["Jun-Ping"],"suffixes":[]},{"propositions":[],"lastnames":["Zhang"],"firstnames":["Chenou"],"suffixes":[]},{"propositions":[],"lastnames":["Renigunta"],"firstnames":["Vijay"],"suffixes":[]},{"propositions":[],"lastnames":["Oliver"],"firstnames":["Dominik"],"suffixes":[]},{"propositions":[],"lastnames":["Navaratnam"],"firstnames":["Dhasakumar","S."],"suffixes":[]},{"propositions":[],"lastnames":["Beckstein"],"firstnames":["Oliver"],"suffixes":[]},{"propositions":[],"lastnames":["Santos-Sacchi"],"firstnames":["Joseph"],"suffixes":[]}],"month":"January","year":"2024","note":"Pages: 2024.01.29.577264 Section: New Results","bibtex":"@misc{bai_chloride_2024,\n\ttitle = {Chloride binding and cholesterol effects on high frequency complex nonlinear capacitance ({cNLC}) in the mouse outer hair cell: experiment and molecular dynamics},\n\tcopyright = {© 2024, Posted by Cold Spring Harbor Laboratory. The copyright holder for this pre-print is the author. All rights reserved. The material may not be redistributed, re-used or adapted without the author's permission.},\n\tshorttitle = {Chloride binding and cholesterol effects on high frequency complex nonlinear capacitance ({cNLC}) in the mouse outer hair cell},\n\turl = {https://www.biorxiv.org/content/10.1101/2024.01.29.577264v1},\n\tdoi = {10.1101/2024.01.29.577264},\n\tabstract = {The function of prestin (SLC26a5), an anion transport family member, has evolved to enhance auditory sensitivity and frequency selectivity by providing mechanical feedback via outer hair cells (OHC) into the organ of Corti. The frequency extent of this boost is governed by the voltage-dependent kinetics of the protein charge movements, otherwise known as nonlinear capacitance (NLC) that we measure in membrane patches under voltage clamp. Here we extend our previous studies on guinea pig OHCs by studying the frequency response of NLC in the mouse OHC, a species with higher frequency auditory needs. We find that the characteristic frequency cut-off (Fis) for the mouse surpasses that of the guinea pig, being 27 kHz vs. 19 kHz, respectively; nevertheless, each shows significant activity in the ultrasonic range. We also evaluate the influence of anion binding on prestin frequency response. Several single point mutations within the chloride binding pocket of prestin (e.g., S396E, S398E) lack anion influence. In agreement, we show absence of anion binding through molecular dynamics (MD) simulations. NLC Fis in the S396E knock-in mouse remains the same as controls, indicating that high frequency activity is likely governed by viscoelastic loads within the membrane characterized by stretched-exponential frequency roll-off. 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