Structural guidance of the photocycle of channelrhodopsin-2 by an interhelical hydrogen bond - PubMed (original) (raw)
. 2010 Jan 19;49(2):267-78.
doi: 10.1021/bi901634p.
Affiliations
- PMID: 20000562
- DOI: 10.1021/bi901634p
Structural guidance of the photocycle of channelrhodopsin-2 by an interhelical hydrogen bond
Christian Bamann et al. Biochemistry. 2010.
Abstract
Channelrhodopsin-2 (ChR2) is a light-gated cation channel and a member of the family of retinylidene photoreceptors. Since the demonstration of light-induced depolarization of ChR2-expressing animal cell membranes, it was increasingly exploited for light triggering of action potentials. ChR2 conducts cations upon light absorption that embodies retinal isomerization as the primary reaction and a structurally unknown opening mechanism. It is evident from spectroscopic data that protonation reactions at the Schiff base are part of the photocycle, comparable to other microbial-type rhodopsins. However, the connection between the processes at the chromophore site and the channel's pore remained enigmatic. Here, we use slow mutants of ChR2 that we generated by disturbing a postulated hydrogen bond when mutating C128 in the transmembrane (TM) helix 3 and D156 in TM helix 4. The lifetime of the mutants' open state is increased more than 100 times. We investigated the spectral properties of the slow mutants. Whereas the deprotonation of the Schiff base (yielding P390) occurs on the same time scale as that of the wild type, reprotonation to P520 is retarded in the slow mutants and their photocycle is split, leading to the presence of two photointermediates, P390 and P520, in the open state. The photoreactions of P390 and P520 lead to a quenching of the current in electrophysiological measurements. We conclude that the putative hydrogen bond between C128 and D156 is an important structural determinant of the channel's closing reaction. Furthermore, we show that the D156A mutant is even more suitable for light control of excitable cells than C128A.
Similar articles
- Spectral characteristics of the photocycle of channelrhodopsin-2 and its implication for channel function.
Bamann C, Kirsch T, Nagel G, Bamberg E. Bamann C, et al. J Mol Biol. 2008 Jan 18;375(3):686-94. doi: 10.1016/j.jmb.2007.10.072. Epub 2007 Nov 1. J Mol Biol. 2008. PMID: 18037436 - The DC gate in Channelrhodopsin-2: crucial hydrogen bonding interaction between C128 and D156.
Nack M, Radu I, Gossing M, Bamann C, Bamberg E, von Mollard GF, Heberle J. Nack M, et al. Photochem Photobiol Sci. 2010 Feb;9(2):194-8. doi: 10.1039/b9pp00157c. Epub 2010 Jan 7. Photochem Photobiol Sci. 2010. PMID: 20126794 - Conformational changes of channelrhodopsin-2.
Radu I, Bamann C, Nack M, Nagel G, Bamberg E, Heberle J. Radu I, et al. J Am Chem Soc. 2009 Jun 3;131(21):7313-9. doi: 10.1021/ja8084274. J Am Chem Soc. 2009. PMID: 19422231 - Channelrhodopsins: directly light-gated cation channels.
Nagel G, Szellas T, Kateriya S, Adeishvili N, Hegemann P, Bamberg E. Nagel G, et al. Biochem Soc Trans. 2005 Aug;33(Pt 4):863-6. doi: 10.1042/BST0330863. Biochem Soc Trans. 2005. PMID: 16042615 Review. - Evolution of the channelrhodopsin photocycle model.
Stehfest K, Hegemann P. Stehfest K, et al. Chemphyschem. 2010 Apr 26;11(6):1120-6. doi: 10.1002/cphc.200900980. Chemphyschem. 2010. PMID: 20349494 Review.
Cited by
- Channel Gating in Kalium Channelrhodopsin Slow Mutants.
Sineshchekov OA, Govorunova EG, Li H, Wang Y, Spudich JL. Sineshchekov OA, et al. J Mol Biol. 2024 Mar 1;436(5):168298. doi: 10.1016/j.jmb.2023.168298. Epub 2023 Oct 5. J Mol Biol. 2024. PMID: 37802216 Free PMC article. - Structural basis for ion selectivity in potassium-selective channelrhodopsins.
Tajima S, Kim YS, Fukuda M, Jo Y, Wang PY, Paggi JM, Inoue M, Byrne EFX, Kishi KE, Nakamura S, Ramakrishnan C, Takaramoto S, Nagata T, Konno M, Sugiura M, Katayama K, Matsui TE, Yamashita K, Kim S, Ikeda H, Kim J, Kandori H, Dror RO, Inoue K, Deisseroth K, Kato HE. Tajima S, et al. Cell. 2023 Sep 28;186(20):4325-4344.e26. doi: 10.1016/j.cell.2023.08.009. Epub 2023 Aug 30. Cell. 2023. PMID: 37652010 Free PMC article. - Optogenetic control of gut movements reveals peristaltic wave-mediated induction of cloacal contractions and reactivation of impaired gut motility.
Shikaya Y, Inaba M, Tadokoro R, Utsunomiya S, Takahashi Y. Shikaya Y, et al. Front Physiol. 2023 May 15;14:1175951. doi: 10.3389/fphys.2023.1175951. eCollection 2023. Front Physiol. 2023. PMID: 37293264 Free PMC article. - Tailoring baker's yeast Saccharomyces cerevisiae for functional testing of channelrhodopsin.
Höler S, Degreif D, Stix F, Yang S, Gao S, Nagel G, Moroni A, Thiel G, Bertl A, Rauh O. Höler S, et al. PLoS One. 2023 Apr 13;18(4):e0280711. doi: 10.1371/journal.pone.0280711. eCollection 2023. PLoS One. 2023. PMID: 37053213 Free PMC article. - The Mechanism of the Channel Opening in Channelrhodopsin-2: A Molecular Dynamics Simulation.
Xin Q, Zhang W, Yuan S. Xin Q, et al. Int J Mol Sci. 2023 Mar 16;24(6):5667. doi: 10.3390/ijms24065667. Int J Mol Sci. 2023. PMID: 36982741 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources