Mechanistic Basis of the Fast Dark Recovery of the Short LOV Protein DsLOV from Dinoroseobacter shibae
| العنوان: | Mechanistic Basis of the Fast Dark Recovery of the Short LOV Protein DsLOV from Dinoroseobacter shibae |
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| المؤلفون: | Ulrich Krauss, Katrin Röllen, Thomas Drepper, Joachim Granzin, Timo Fettweiss, Stephan Endres, Oliver Reiners, Dieter Willbold, Karl-Erich Jaeger, Renu Batra-Safferling |
| المصدر: | Biochemistry. 57:4833-4847 |
| بيانات النشر: | American Chemical Society (ACS), 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 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 |
| الوصف: | 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. |
| تدمد: | 1520-4995 0006-2960 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::102a8b3c8601993a3ae36218e4d18784Test https://doi.org/10.1021/acs.biochem.8b00645Test |
| رقم الانضمام: | edsair.doi.dedup.....102a8b3c8601993a3ae36218e4d18784 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15204995 00062960 |
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