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@ARTICLE{Remeeva:904327,
author = {Remeeva, Alina and Nazarenko, Vera V. and Kovalev, Kirill
and Goncharov, Ivan M. and Yudenko, Anna and Astashkin,
Roman and Gordeliy, Valentin and Gushchin, Ivan},
title = {{I}nsights into the mechanisms of light‐oxygen‐voltage
domain color tuning from a set of high‐resolution
{X}‐ray structures},
journal = {Proteins},
volume = {89},
number = {8},
issn = {0887-3585},
address = {New York, NY},
publisher = {Wiley-Liss},
reportid = {FZJ-2021-05897},
pages = {1005 - 1016},
year = {2021},
abstract = {Light-oxygen-voltage (LOV) domains are widespread
photosensory modules that can be used in fluorescence
microscopy, optogenetics and controlled production of
reactive oxygen species. All of the currently known LOV
domains have absorption maxima in the range of ~440 to
~450 nm, and it is not clear whether they can be shifted
significantly using mutations. Here, we have generated a
panel of LOV domain variants by mutating the key
chromophore-proximal glutamine aminoacid of a thermostable
flavin based fluorescent protein CagFbFP (Gln148) to
asparagine, aspartate, glutamate, histidine, lysine and
arginine. Absorption spectra of all of the mutants are
blue-shifted, with the maximal shift of 8 nm observed for
the Q148H variant. While CagFbFP and its Q148N/D/E variants
are not sensitive to pH, Q148H/K/R reveal a moderate red
shift induced byacidic pH. To gain further insight, we
determined high resolution crystal structures of all of the
mutants studied at the resolutions from 1.07 Å for Q148D
to 1.63 Å for Q148R. Whereas in some of the variants, the
aminoacid 148 remains in the vicinity of the flavin, in
Q148K, Q148R and partially Q148D, the C-terminus of the
protein unlatches and the side chain of the residue 148 is
reoriented away from the chromophore. Our results explain
the absence of color shifts from replacing Gln148 with
charged aminoacids and pave the way for rational design of
color-shifted flavin based fluorescent proteins.},
cin = {IBI-7},
ddc = {570},
cid = {I:(DE-Juel1)IBI-7-20200312},
pnm = {5241 - Molecular Information Processing in Cellular Systems
(POF4-524)},
pid = {G:(DE-HGF)POF4-5241},
typ = {PUB:(DE-HGF)16},
pubmed = {33774867},
UT = {WOS:000635931200001},
doi = {10.1002/prot.26078},
url = {https://juser.fz-juelich.de/record/904327},
}