Channelrhodopsin C1C2: Photocycle kinetics and interactions near the central gate
Abstract
Channelrhodopsins (ChR) are light-sensitive cation channels used in optogenetics, a technique that applies light to control cells (e.g., neurons) that have been modified genetically to express those channels. Although mutations are known to affect pore kinetics, little is known about how mutations induce changes at the molecular scale. To address this issue, we first measured channel opening and closing rates of a ChR chimera (C1C2) and selected variants (N297D, N297V, and V125L). Then, we used atomistic simulations to correlate those rates with changes in pore structure, hydration, and chemical interactions among key gating residues of C1C2 in both closed and open states. Overall, the experimental results show that C1C2 and its mutants do not behave like ChR2 or its analogous variants, except V125L, making C1C2 a unique channel. Furthermore, our atomistic simulations confirmed that opening of the channel and initial hydration of the gating regions between helices I, II, III, and VII of the channel occurs with 1) the presence of 13-cis retinal; 2) deprotonation of a glutamic acid gating residue, E129; and 3) subsequent weakening of the central gate hydrogen bond between the same glutamic acid E129 and asparagine N297 in the central region of the pore. Also, anmore »
- Authors:
-
[2];
[2];
[3]
- Univ. of New Orleans, LA (United States)
- Worcester Polytechnic Inst., MA (United States)
- Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
- Publication Date:
- Research Org.:
- Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
- Sponsoring Org.:
- USDOE National Nuclear Security Administration (NNSA)
- OSTI Identifier:
- 1778036
- Report Number(s):
- SAND-2021-3722J
Journal ID: ISSN 0006-3495; 695097
- Grant/Contract Number:
- AC04-94AL85000; NA0003525
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Biophysical Journal
- Additional Journal Information:
- Journal Volume: 120; Journal Issue: 9; Journal ID: ISSN 0006-3495
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
VanGordon, Monika R., Prignano, Lindsey A., Dempski, Robert E., Rick, Steven W., and Rempe, Susan B. Channelrhodopsin C1C2: Photocycle kinetics and interactions near the central gate. United States: N. p., 2021.
Web. doi:10.1016/j.bpj.2021.03.002.
VanGordon, Monika R., Prignano, Lindsey A., Dempski, Robert E., Rick, Steven W., & Rempe, Susan B. Channelrhodopsin C1C2: Photocycle kinetics and interactions near the central gate. United States. https://doi.org/10.1016/j.bpj.2021.03.002
VanGordon, Monika R., Prignano, Lindsey A., Dempski, Robert E., Rick, Steven W., and Rempe, Susan B. Tue .
"Channelrhodopsin C1C2: Photocycle kinetics and interactions near the central gate". United States. https://doi.org/10.1016/j.bpj.2021.03.002. https://www.osti.gov/servlets/purl/1778036.
@article{osti_1778036,
title = {Channelrhodopsin C1C2: Photocycle kinetics and interactions near the central gate},
author = {VanGordon, Monika R. and Prignano, Lindsey A. and Dempski, Robert E. and Rick, Steven W. and Rempe, Susan B.},
abstractNote = {Channelrhodopsins (ChR) are light-sensitive cation channels used in optogenetics, a technique that applies light to control cells (e.g., neurons) that have been modified genetically to express those channels. Although mutations are known to affect pore kinetics, little is known about how mutations induce changes at the molecular scale. To address this issue, we first measured channel opening and closing rates of a ChR chimera (C1C2) and selected variants (N297D, N297V, and V125L). Then, we used atomistic simulations to correlate those rates with changes in pore structure, hydration, and chemical interactions among key gating residues of C1C2 in both closed and open states. Overall, the experimental results show that C1C2 and its mutants do not behave like ChR2 or its analogous variants, except V125L, making C1C2 a unique channel. Furthermore, our atomistic simulations confirmed that opening of the channel and initial hydration of the gating regions between helices I, II, III, and VII of the channel occurs with 1) the presence of 13-cis retinal; 2) deprotonation of a glutamic acid gating residue, E129; and 3) subsequent weakening of the central gate hydrogen bond between the same glutamic acid E129 and asparagine N297 in the central region of the pore. Also, an aspartate (D292) is the unambiguous primary proton acceptor for the retinal Schiff base in the hydrated channel.},
doi = {10.1016/j.bpj.2021.03.002},
journal = {Biophysical Journal},
number = 9,
volume = 120,
place = {United States},
year = {Tue Mar 09 00:00:00 EST 2021},
month = {Tue Mar 09 00:00:00 EST 2021}
}
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