Journal
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
Volume 140, Issue 31, Pages 9899-9903Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jacs.8b03931
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Funding
- Spanish Ministry of Economy, Industry and Competitiveness [BFU2016-768050-P, RYC-2013-13114]
- German Research Foundation [SFB-1078]
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Channelrhodopsins (ChRs) are light-gated cation channels. In spite of their wide use to activate neurons with light, the photocurrents of ChRs rapidly decay in intensity under both continuous illumination and fast trains of light pulses, broadly referred to as desensitization. This undesirable phenomenon has been explained by two interconnected photocycles, each of them containing a nonconductive dark state (D1 and D2) and a conductive state (O1 and O2). While the D1 and O1 states correspond to the dark-state and P-3(520) intermediate of the primary all-trans photocycle of ChR2, the molecular identity of D2 and O2 remains unclear. We show that P-4(480), the last intermediate of the all-trans photocycle, is photoactive. Its photocycle, characterized by time-resolved UV/vis spectroscopy, contains a red-shifted intermediate, I-3(530). Our results indicate that the D2 and O2 states correspond to the P-4(480) and I-3(530) intermediates, connecting desensitization of ChR2 with the photochemical properties of the P-4(480) intermediate.
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