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@ARTICLE{Nazarenko:866224,
author = {Nazarenko, Vera V. and Remeeva, Alina and Yudenko, Anna and
Kovalev, Kirill and Dubenko, Anton and Goncharov, Ivan M.
and Kuzmichev, Pavel and Rogachev, Andrey V. and Buslaev,
Pavel and Borshchevskiy, Valentin and Mishin, Alexey and
Dhoke, Gaurao V. and Schwaneberg, Ulrich and Davari, Mehdi
D. and Jaeger, Karl-Erich and Krauss, Ulrich and Gordeliy,
Valentin and Gushchin, Ivan},
title = {{A} thermostable flavin-based fluorescent protein from
{C}hloroflexus aggregans : a framework for ultra-high
resolution structural studies},
journal = {Photochemical $\&$ photobiological sciences},
volume = {18},
number = {7},
issn = {1474-9092},
address = {Cambridge},
publisher = {Royal Society of Chemistry},
reportid = {FZJ-2019-05390},
pages = {1793 - 1805},
year = {2019},
abstract = {Light-Oxygen-Voltage (LOV) domains are conserved parts of
photoreceptors in plants, bacteria and fungi that bind
flavins as chromophores and detect blue light. In the past,
LOV domain variants have been developed as fluorescent
reporter proteins (called flavin-based fluorescent proteins;
FbFPs), which due to their ability to fluoresce under
anaerobic conditions, fast folding kinetics and a small size
of ∼12–16 kDa are a promising reporter system for
quantitative real-time analysis of biological processes.
Here, we present a small thermostable flavin-based
fluorescent protein CagFbFP derived from a soluble LOV
domain-containing histidine kinase from the thermophilic
bacterium Chloroflexus aggregans. CagFbFP is composed of 107
amino acids with a molecular weight of 11.6 kDa and consists
only of the conserved LOV core domain. The protein is
thermostable with a melting point of about 68 °C. It
crystallizes easily and its crystals diffract to 1.07 Å.
Both the crystal structure and small angle scattering data
show that the protein is a dimer. Unexpectedly, glutamine
148, which in LOV photoreceptor proteins is the key residue
responsible for signal transduction, occupies two
conformations. Molecular dynamics simulations show that the
two conformations interconvert rapidly. The crystal
structure of the wild-type Chloroflexus aggregans LOV domain
determined at 1.22 Å resolution confirmed the presence of
two alternative conformations of the glutamine 148 side
chain. Overall, this protein, due to its stability and ease
of crystallization, appears to be a promising model for
ultra-high resolution structural studies of LOV domains and
for application as a fluorescent reporter.},
cin = {IMET / ICS-6 / IBG-1},
ddc = {620},
cid = {I:(DE-Juel1)IMET-20090612 / I:(DE-Juel1)ICS-6-20110106 /
I:(DE-Juel1)IBG-1-20101118},
pnm = {581 - Biotechnology (POF3-581)},
pid = {G:(DE-HGF)POF3-581},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:31116222},
UT = {WOS:000477947100017},
doi = {10.1039/C9PP00067D},
url = {https://juser.fz-juelich.de/record/866224},
}