TY  - JOUR
AU  - Fettweiss, Timo
AU  - Röllen, Katrin
AU  - Granzin, Joachim
AU  - Reiners, Oliver
AU  - Endres, Stephan
AU  - Drepper, Thomas
AU  - Willbold, Dieter
AU  - Jaeger, Karl-Erich
AU  - Batra-Safferling, Renu
AU  - Krauss, Ulrich
TI  - Mechanistic Basis of the Fast Dark Recovery of the Short LOV Protein DsLOV from Dinoroseobacter shibae
JO  - Biochemistry
VL  - 57
IS  - 32
SN  - 1520-4995
CY  - Columbus, Ohio
PB  - American Chemical Society
M1  - FZJ-2018-04636
SP  - 4833–4847
PY  - 2018
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - pmid:29989797
UR  - <Go to ISI:>//WOS:000442184600008
DO  - DOI:10.1021/acs.biochem.8b00645
UR  - https://juser.fz-juelich.de/record/850885
ER  -