Tracking Pore Hydration in Channelrhodopsin by Site-Directed Infrared-Active Azido Probes
| Autor: | Franz Bartl, Jens Kuhne, Joel C.D. Kaufmann, Johannes Vierock, Klaus Gerwert, Thomas Huber, Thomas P. Sakmar, Peter Hegemann, Benjamin S. Krause |
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| Rok vydání: | 2019 |
| Předmět: |
Azides
0303 health sciences Chemistry Infrared Phenylalanine fungi 030302 biochemistry & molecular biology Channelrhodopsin Gating Tracking (particle physics) Biochemistry 03 medical and health sciences HEK293 Cells Channelrhodopsins Molecular Probes Spectroscopy Fourier Transform Infrared Codon Terminator Mutagenesis Site-Directed Biophysics Humans |
| Zdroj: | Biochemistry. 58:1275-1286 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/acs.biochem.8b01211 |
| Popis: | In recent years, gating and transient ion-pathway formation in the light-gated channelrhodopsins (ChRs) have been intensively studied. Despite these efforts, a profound understanding of the mechanistic details is still lacking. To track structural changes concomitant with the formation and subsequent collapse of the ion-conducting pore, we site-specifically introduced the artificial polarity-sensing probe p-azido-l-phenylalanine (azF) into several ChRs by amber stop codon suppression. The frequently used optogenetic actuator ReaChR (red-activatable ChR) exhibited the best expression properties of the wild type and the azF mutants. By exploiting the unique infrared spectral absorption of azF [ν |
| Databáze: | OpenAIRE |
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