Mechanistic Basis of the Fast Dark Recovery of the Short LOV Protein DsLOV from Dinoroseobacter shibae
| Autor: | Ulrich Krauss, Katrin Röllen, Thomas Drepper, Joachim Granzin, Timo Fettweiss, Stephan Endres, Oliver Reiners, Dieter Willbold, Karl-Erich Jaeger, Renu Batra-Safferling |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Light genetic structures Protein Conformation Optogenetics Crystallography X-Ray Photoreceptors Microbial 010402 general chemistry medicine.disease_cause 01 natural sciences Biochemistry 03 medical and health sciences Protein structure Bacterial Proteins medicine Rhodobacteraceae Phototropism Mutation Pseudomonas putida Chemistry Circular Dichroism Mutagenesis Dinoroseobacter shibae 0104 chemical sciences Oxygen Kinetics 030104 developmental biology Dark state Mutagenesis Site-Directed Biophysics sense organs Function (biology) Transcription Factors |
| Zdroj: | Biochemistry. 57:4833-4847 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/acs.biochem.8b00645 |
| Popis: | Light, oxygen, voltage (LOV) proteins, a ubiquitously distributed class of photoreceptors, regulate a wide variety of light-dependent physiological responses. Because of their modular architecture, LOV domains, i.e., the sensory domains of LOV photoreceptors, have been widely used for the construction of optogenetic tools. We recently described the structure and function of a short LOV protein (DsLOV) from the marine phototropic bacterium Dinoroseobacter shibae, for which, in contrast to other LOV photoreceptors, the dark state represents the physiologically relevant signaling state. Among bacterial LOV photoreceptors, DsLOV possesses an exceptionally fast dark recovery, corroborating its function as a "dark" sensor. To address the mechanistic basis of this unusual characteristic, we performed a comprehensive mutational, kinetic, thermodynamic, and structural characterization of DsLOV. The mechanistic basis of the fast dark recovery of the protein was revealed by mutation of the previously noted uncommon residue substitution at position 49 found in DsLOV. The substitution of M49 with different residues that are naturally conserved in LOV domains tuned the dark-recovery time of DsLOV over 3 orders of magnitude, without grossly affecting its overall structure or the light-dependent structural change observed for the wild-type protein. Our study thus provides a striking example of how nature can achieve LOV photocycle tuning by subtle structural alterations in the LOV domain active site, highlighting the easy evolutionary adaptability of the light sensory function. At the same time, our data provide guidance for the mutational photocycle tuning of LOV domains, with relevance for the growing field of optogenetics. |
| Databáze: | OpenAIRE |
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