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@ARTICLE{LaheyRudolph:450351,
author = {Lahey-Rudolph, Janine Mia and Schönherr, Robert and
Jeffries, Cy M. and Blanchet, Clément E. and Boger, Juliane
and Ferreira Ramos, Ana Sofia and Riekehr, Winnie Maria and
Triandafillidis, Dimitris-Panagiotis and Valmas, Alexandros
and Margiolaki, Irene and Svergun, Dmitri and Redecke, Lars},
title = {{R}apid screening of in cellulo grown protein crystals via
a small-angle {X}-ray scattering/{X}-ray powder diffraction
synergistic approach},
journal = {Journal of applied crystallography},
volume = {53},
number = {5},
issn = {1600-5767},
address = {[S.l.]},
publisher = {Wiley-Blackwell},
reportid = {PUBDB-2020-04297},
pages = {1169 - 1180},
year = {2020},
abstract = {Crystallization of recombinant proteins in living cells is
an exciting new approach for structural biology that
provides an alternative to the time-consuming optimization
of protein purification and extensive crystal screening
steps. Exploiting the potential of this approach requires a
more detailed understanding of the cellular processes
involved and versatile screening strategies for crystals in
a cell culture. Particularly if the target protein forms
crystalline structures of unknown morphology only in a small
fraction of cells, their detection by applying standard
visualization techniques can be time consuming and difficult
owing to the environmental challenges imposed by the living
cells. In this study, a high-brilliance and low-background
bioSAXS beamline is employed for rapid and sensitive
detection of protein microcrystals grown within insect
cells. On the basis of the presence of Bragg peaks in the
recorded small-angle X-ray scattering profiles, it is
possible to assess within seconds whether a cell culture
contains microcrystals, even in a small percentage of cells.
Since such information cannot be obtained by other
established detection methods in this time frame, this
screening approach has the potential to overcome one of the
bottlenecks of intracellular crystal detection. Moreover,
the association of the Bragg peak positions in the
scattering curves with the unit-cell composition of the
protein crystals raises the possibility of investigating the
impact of environmental conditions on the crystal structure
of the intracellular protein crystals. This information
provides valuable insights helping to further understand the
in cellulo crystallization process.},
cin = {EMBL-User / EMBL / FS-CFEL-1 / DOOR ; HAS-User / U Lübeck},
ddc = {540},
cid = {I:(DE-H253)EMBL-User-20120814 / I:(DE-H253)EMBL-20120731 /
I:(DE-H253)FS-CFEL-1-20120731 / I:(DE-H253)HAS-User-20120731
/ $I:(DE-H253)U_L__beck-20211012$},
pnm = {633 - Life Sciences – Building Blocks of Life: Structure
and Function (POF4-633) / 6G3 - PETRA III (POF3-622)},
pid = {G:(DE-HGF)POF4-633 / G:(DE-HGF)POF3-6G3},
experiment = {EXP:(DE-H253)P-P12-20150101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:33117106},
UT = {WOS:000577178000001},
doi = {10.1107/S1600576720010687},
url = {https://bib-pubdb1.desy.de/record/450351},
}