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Title: Channelrhodopsin-2 Function is Modulated by Residual Hydrophobic Mismatch with the Surrounding Lipid Environment

Abstract

Channelrhodopsin-2 (ChR2) is a light-gated ion channel that conducts cations of multiple valencies down the electrochemical gradient. This light-gated property has made ChR2 a popular tool in the field of optogenetics, allowing for the spatial and temporal control of excitable cells with light. A central aspect of protein function is the interaction with the surrounding lipid environment. To further explore these membrane-protein interactions, we demonstrate the role of residual hydrophobic mismatch (RHM) as a mechanistically important component of ChR2 function. We combined computational and functional experiments to understand how RHM between the lipid environment and ChR2 alters the structural and biophysical properties of the channel. Analysis of our results revealed significant RHM at the intracellular/lipid interface of ChR2 from a triad of residues. The resulting energy penalty is substantial and can be lowered via mutagenesis to evaluate the functional effects of this change in lipid-protein interaction energy. The experimental measurement of channel stability, conductance and selectivity resulting from the reduction of the RHM energy penalty showed changes in progressive H+ permeability, kinetics and open-state stability, suggesting how the modulation of ChR2 by the surrounding lipid membrane can play an important biological role and contribute to the design of targeted optogeneticmore » constructs for specific cell types.« less

Authors:
; ; ORCiD logo;
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1530465
Grant/Contract Number:  
AC02- 05CH11231
Resource Type:
Published Article
Journal Name:
Applied Sciences
Additional Journal Information:
Journal Name: Applied Sciences Journal Volume: 9 Journal Issue: 13; Journal ID: ISSN 2076-3417
Publisher:
MDPI AG
Country of Publication:
Switzerland
Language:
English

Citation Formats

Richards, Ryan, Mondal, Sayan, Weinstein, Harel, and Dempski, Robert E. Channelrhodopsin-2 Function is Modulated by Residual Hydrophobic Mismatch with the Surrounding Lipid Environment. Switzerland: N. p., 2019. Web. doi:10.3390/app9132674.
Richards, Ryan, Mondal, Sayan, Weinstein, Harel, & Dempski, Robert E. Channelrhodopsin-2 Function is Modulated by Residual Hydrophobic Mismatch with the Surrounding Lipid Environment. Switzerland. doi:10.3390/app9132674.
Richards, Ryan, Mondal, Sayan, Weinstein, Harel, and Dempski, Robert E. Sun . "Channelrhodopsin-2 Function is Modulated by Residual Hydrophobic Mismatch with the Surrounding Lipid Environment". Switzerland. doi:10.3390/app9132674.
@article{osti_1530465,
title = {Channelrhodopsin-2 Function is Modulated by Residual Hydrophobic Mismatch with the Surrounding Lipid Environment},
author = {Richards, Ryan and Mondal, Sayan and Weinstein, Harel and Dempski, Robert E.},
abstractNote = {Channelrhodopsin-2 (ChR2) is a light-gated ion channel that conducts cations of multiple valencies down the electrochemical gradient. This light-gated property has made ChR2 a popular tool in the field of optogenetics, allowing for the spatial and temporal control of excitable cells with light. A central aspect of protein function is the interaction with the surrounding lipid environment. To further explore these membrane-protein interactions, we demonstrate the role of residual hydrophobic mismatch (RHM) as a mechanistically important component of ChR2 function. We combined computational and functional experiments to understand how RHM between the lipid environment and ChR2 alters the structural and biophysical properties of the channel. Analysis of our results revealed significant RHM at the intracellular/lipid interface of ChR2 from a triad of residues. The resulting energy penalty is substantial and can be lowered via mutagenesis to evaluate the functional effects of this change in lipid-protein interaction energy. The experimental measurement of channel stability, conductance and selectivity resulting from the reduction of the RHM energy penalty showed changes in progressive H+ permeability, kinetics and open-state stability, suggesting how the modulation of ChR2 by the surrounding lipid membrane can play an important biological role and contribute to the design of targeted optogenetic constructs for specific cell types.},
doi = {10.3390/app9132674},
journal = {Applied Sciences},
number = 13,
volume = 9,
place = {Switzerland},
year = {2019},
month = {6}
}

Journal Article:
Free Publicly Available Full Text
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DOI: 10.3390/app9132674

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