| Author/Editor | Gourinchas, Geoffrey; Vide, Uršula; Winkler, Andreas | |
| Title | Influence of the N-terminal segment and the PHY-tongue element on light-regulation in bacteriophytochromes | |
| Type | članek | |
| Publication year | 2019 | |
| Volume | str. 1-20 | |
| ISSN | 1083-351X - The Journal of biological chemistry | |
| Language | eng | |
| Abstract | Photoreceptors enable the integration of ambient light stimuli to trigger lifestyle adaptations via modulation of central metabolite levels involved in diverse regulatory processes. Red light sensing bacteriophytochromes are attractive targets for the development of innovative optogenetic tools due to their natural modularity of coupling with diverse functionalities and the natural availability of the light-absorbing biliverdin chromophore in animal tissues. However, a rational design of such tools is complicated by the poor understanding of molecular mechanisms of light signal transduction over long distances - from the site of photon absorption to the active site of downstream enzymatic effectors. Here we show how swapping structural elements between two bacteriophytochrome homologs provides additional insights into light signal integration and effector regulation, involving a fine-tuned interplay of important structural elements of the sensor as well as the sensor-effector linker. Facilitated by the availability of structural information of inhibited and activated full-length structures of one of the two homologs (Idiomarina species A28L Phytochrome activated diguanylyl Cyclase - IsPadC) and characteristic differences in photoresponses of the two homologs, we identify an important cross-talk between the N-terminal segment (NTS), containing the covalent attachment site of the chromophore, and the PHY-tongue region. Moreover, we highlight how these elements influence the dynamic range of photoactivation and how activation can be improved to light/dark ratios of ~800-fold by reducing basal dark-state activities at the same time as increasing conversion in the light-state. This will enable future optimization of optogenetic tools aiming at a direct allosteric regulation of enzymatic effectors. | |
| Keywords | phytochrome ultraviolet-visible spectroscopy (UV-Vis spectroscopy) photobiology fitokrom ultravijolična vidna spektroskopija (UV-Vis spektroskopija) fotobiologija |