| Autor: |
Hisashi Naitow, Ayumi Yamashita, Michihiro Sugahara, Mikako Shirouzu, Rie Tanaka, Tamao Hisano, Shun-ichi Sekine, Haruhiko Ehara, Takashi Nomura, Kentaro Ihara, Yoshinori Matsuura, Tomoyuki Tanaka, Shigeki Owada, Takuhiro Ito, Osamu Nureki, Ryogo Akasaka, Kazushige Katsura, Kazutaka Murayama, Susumu Yoshizawa, Tomomi Kimura-Someya, Kazuhiro Kashiwagi, Toshiaki Hosaka, Toshi Arima, Luo Fangjia, So Iwata, Eriko Nango, Takanori Nakane, Minoru Kubo, Kensuke Tono |
| Rok vydání: |
2021 |
| Předmět: |
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| Popis: |
Light-driven chloride-pumping rhodopsins actively transport anions, including various halide ions, across cell membranes. Recent studies using time-resolved serial femtosecond crystallography (TR-SFX) have uncovered the structural changes and ion transfer mechanisms in light-driven cation-pumping rhodopsins. However, the mechanism by which the conformational changes pump an anion to achieve unidirectional ion transport, from the extracellular side to the cytoplasmic side, in anion-pumping rhodopsins remains enigmatic. We have collected TR-SFX data ofNonlabens marinusrhodopsin-3 (NM-R3), derived from a marine flavobacterium, at 10 μs and 1 ms time-points after photoexcitation. Our structural analysis reveals the conformational alterations during ion transfer and after ion release. Movements of the retinal chromophore initially displace a conserved tryptophan to the cytoplasmic side of NM-R3, accompanied with a slight shift of the halide ion bound to the retinal. After ion release, the inward movements of helix C and helix G and the lateral displacements of the retinal block access to the extracellular side of NM-R3. Anomalous signal data have also been obtained from NM-R3 crystals containing iodide ions. The anomalous density maps provide insight into the halide binding site for ion transfer in NM-R3.SignificanceLight-driven chloride pumps have been identified in various species, including archaea and marine flavobacteria. The function of ion transportation controllable by light is utilized for optogenetics tools in neuroscience. Chloride pumps differ among species, in terms of amino acid homology and structural similarity. Our time-resolved crystallographic studies using X-ray free electron lasers reveal the molecular mechanism of halide ion transfer in a light-driven chloride pump from a marine flavobacterium. Our data indicate a common mechanism in chloride pumping rhodopsins, as compared to previous low temperature trapping studies of chloride pumps. These findings are significant not only for further improvements of optogenetic tools but also for a general understanding of the ion pumping mechanisms of microbial rhodopsins. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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