TY  - JOUR
A1  - Becker-Baldus, Johanna
A1  - Leeder, Alexander
A1  - Brown, Lynda J.
A1  - Brown, C. D.
A1  - Bamann, Christian
A1  - Glaubitz, Clemens
T1  - The desensitized channelrhodopsin-2 photointermediate contains 13 -cis, 15 -syn retinal Schiff base
T2  - Angewandte Chemie
N2  - Channelrhodopsin-2 (ChR2) is a light-gated cation channel and was used to lay the foundations of optogenetics. Its dark state X-ray structure has been determined in 2017 for the wild-type, which is the prototype for all other ChR variants. However, the mechanistic understanding of the channel function is still incomplete in terms of structural changes after photon absorption by the retinal chromophore and in the framework of functional models. Hence, detailed information needs to be collected on the dark state as well as on the different photointermediates. For ChR2 detailed knowledge on the chromophore configuration in the different states is still missing and a consensus has not been achieved. Using DNP-enhanced solid-state MAS NMR spectroscopy on proteoliposome samples, we unambiguously determined the chromophore configuration in the desensitized state, and we show that this state occurs towards the end of the photocycle.
KW  - channelrhodopsin
KW  - dynamic nuclear polarization
KW  - membrane proteins
KW  - photocycle
KW  - solid-state NMR spectroscopy
Y1  - 2021
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/63892
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-638929
SN  - 1521-3773
N1  - The work was funded by Deutsche Forschungs-gemeinschaft/Sonderforschungsbereich 807 Transport and Communications across Membranes. The dynamic nuclear polarization experiments were enabled through DFG Equipment Grant GL 307/4-1 and the Cluster of Excellence Frankfurt: Macromolecular Complexes Frankfurt. Work at the Center for Biomolecular Magnetic Resonance is supported by the State of Hesse. Open access funding enabled and organized by Projekt DEAL.
VL  - 60
IS  - 30
SP  - 16442
EP  - 16447
PB  - Wiley-VCH
CY  - Weinheim
ER  -