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@ARTICLE{Zabelskii:626502,
author = {Zabelskii, Dmitrii and Bukhdruker, Sergey and Bukhalovich,
Siarhei and Tsybrov, Fedor and Lamm, Gerrit H. U. and
Astashkin, Roman and Doroginin, Demid and Matveev, Grigory
and Sudarev, Vsevolod and Kuzmin, Alexander and Zinovev,
Egor and Vlasova, Anastasiia and Ryzhykau, Yury and
Ilyinsky, Nikolay and Gushchin, Ivan and Bourenkov, Gleb and
Alekseev, Alexey and Round, Adam and Wachtveitl, Josef and
Bamberg, Ernst and Gordeliy, Valentin},
title = {{I}on-conducting and gating molecular mechanisms of
channelrhodopsin revealed by true-atomic-resolution
structures of open and closed states},
journal = {Nature structural $\&$ molecular biology},
volume = {32},
number = {8},
issn = {1545-9993},
address = {London [u.a.]},
publisher = {Nature Publishing Group},
reportid = {PUBDB-2025-01455},
pages = {1347 - 1357},
year = {2025},
note = {Waiting for fulltext},
abstract = {Channelrhodopsins (ChRs) have emerged as major optogenetics
tools, particularly in neuroscience. Despite their
importance, the molecular mechanism of ChR opening remains
elusive. Moreover, all reported structures of ChRs
correspond to either a closed or an early intermediate state
and lack the necessary level of detail owing to the limited
resolution. Here we present the structures of the closed and
open states of a cation-conducting ChR, OLPVR1, from Organic
Lake phycodnavirus, belonging to the family of viral ChRs
solved at 1.1- and 1.3-Å resolution at physiologically
relevant pH conditions (pH 8.0). OLPVR1 was expressed in
Escherichia coli and crystallized using an in meso approach,
and the structures were solved by X-ray crystallography. We
also present the structure of the OLPVR1 protonated state at
acidic pH (pH 2.5) at 1.4-Å resolution. Together, these
three structures elucidate the molecular mechanisms of the
channel’s opening and permeability in detail. Extensive
functional studies support the proposed mechanisms. Channel
opening is controlled by isomerization of the retinal
cofactor, triggering protonation of proton acceptors and
deprotonation of proton donors located in the three gates of
the channel. The E51 residue in the core of the central gate
(similar to E90 of ChR2 from Chlamydomonas reinhardtii)
plays a key role in the opening of the channel. E51 flips
out of the gate and towards the proton acceptor D200 (D253
in ChR2 in C. reinhardtii), establishing a hydrogen bond
between them. Despite differences in subfamilies of ChRs,
they share a common gate–cavity architecture, suggesting
that they could have similar general gating mechanisms.
These results enabled us to design viral rhodopsin with
improved properties for optogenetic applications. The
structural data and mechanisms might also be helpful for
better understanding other ChRs and their engineering.},
cin = {EMBL-User / EMBL},
ddc = {570},
cid = {I:(DE-H253)EMBL-User-20120814 / I:(DE-H253)EMBL-20120731},
pnm = {6G3 - PETRA III (DESY) (POF4-6G3)},
pid = {G:(DE-HGF)POF4-6G3},
experiment = {EXP:(DE-H253)P-P14-20150101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:40205223},
doi = {10.1038/s41594-025-01488-7},
url = {https://bib-pubdb1.desy.de/record/626502},
}