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@ARTICLE{Golub:873376,
author = {Golub, Maksym and Moldenhauer, Marcus and Schmitt,
Franz-Josef and Feoktystov, Artem and Mändar, Hugo and
Maksimov, Eugene and Friedrich, Thomas and Pieper, Jörg},
title = {{S}olution {S}tructure and {C}onformational {F}lexibility
in the {A}ctive {S}tate of the {O}range {C}arotenoid
{P}rotein: {P}art {I}. {S}mall-{A}ngle {S}cattering},
journal = {The journal of physical chemistry / B B, Condensed matter,
materials, surfaces, interfaces $\&$ biophysical},
volume = {123},
number = {45},
issn = {1520-5207},
address = {Washington, DC},
publisher = {Soc.},
reportid = {FZJ-2020-00688},
pages = {9525 - 9535},
year = {2019},
abstract = {Orange carotenoid proteins (OCPs) are photoswitchable
macromolecules playing an important role in nonphotochemical
quenching of excess energy in cyanobacterial light
harvesting. Upon absorption of a blue photon (450–500 nm),
OCPs undergo a structural change from the ground state OCPO
to the active state OCPR, but high-resolution structures of
the active state OCPR are not yet available. Here, we use
small-angle scattering methods combined with simulation
tools to determine low-resolution structures of the active
state at low protein concentrations via two approaches:
first, directly by in situ illumination of wild-type OCP
achieving a turnover to the active state of $>90\%$ and
second, by using the mutant OCPW288A anticipated to mimic
the active state structure. Data fits assuming the shape of
an ellipsoid yield three ellipsoidal radii of about 9, 29,
and 51 ± 1 Å, in the case of the ground state OCPO. In the
active state, however, the molecule becomes somewhat
narrower with the two smaller radii being 9 and only 19 ± 3
Å, while the third dimension of the ellipsoid is
significantly elongated to 85–92 ± 5 Å. Reconstitutions
of the active state structure corroborate that OCPR is
significantly elongated compared to the ground state OCPO
and characterized by a separation of the N-terminal and
C-terminal domains with unfolded N-terminal extension. By
direct comparison of small-angle scattering data, we
directly show that the mutant OCPW288A can be used as a
structural analogue of the active state OCPR. The
small-angle experiments are repeated for OCPO and the mutant
OCPW288A at high protein concentrations of 50–65 mg/mL
required for neutron spectroscopy investigating the
molecular dynamics of OCP (see accompanying paper). The
results reveal that the OCPO and OCPW288A samples for
dynamics experiments are preferentially dimeric and widely
resemble the structures of the ground and active states of
OCP, respectively. This enables us to properly characterize
the molecular dynamics of both states of OCP in the
accompanying paper.},
cin = {JCNS-FRM-II / JCNS-2 / MLZ},
ddc = {530},
cid = {I:(DE-Juel1)JCNS-FRM-II-20110218 /
I:(DE-Juel1)JCNS-2-20110106 / I:(DE-588b)4597118-3},
pnm = {6G4 - Jülich Centre for Neutron Research (JCNS) (POF3-623)
/ 6G15 - FRM II / MLZ (POF3-6G15)},
pid = {G:(DE-HGF)POF3-6G4 / G:(DE-HGF)POF3-6G15},
experiment = {EXP:(DE-MLZ)KWS1-20140101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:31556613},
UT = {WOS:000497259800002},
doi = {10.1021/acs.jpcb.9b05071},
url = {https://juser.fz-juelich.de/record/873376},
}