TY  - JOUR
AU  - Schönherr, Robert
AU  - Boger, Juliane
AU  - Lahey-Rudolph, J. Mia
AU  - Harms, Mareike
AU  - Kaiser, Jacqueline
AU  - Nachtschatt, Sophie
AU  - Wobbe, Marla
AU  - Duden, Rainer
AU  - König, Peter
AU  - Bourenkov, Gleb
AU  - Schneider, Thomas R.
AU  - Redecke, Lars
TI  - A streamlined approach to structure elucidation using in cellulo crystallized recombinant proteins, InCellCryst
JO  - Nature Communications
VL  - 15
IS  - 1
SN  - 2041-1723
CY  - [London]
PB  - Nature Publishing Group UK
M1  - PUBDB-2024-01564
SP  - 1709
PY  - 2024
AB  - With the advent of serial X-ray crystallography on microfocus beamlines at free-electron laser and synchrotron facilities, the demand for protein microcrystals has significantly risen in recent years. However, by in vitro crystallization extensive efforts are usually required to purify proteins and produce sufficiently homogeneous microcrystals. Here, we present InCellCryst, an advanced pipeline for producing homogeneous microcrystals directly within living insect cells. Our baculovirus-based cloning system enables the production of crystals from completely native proteins as well as the screening of different cellular compartments to maximize chances for protein crystallization. By optimizing cloning procedures, recombinant virus production, crystallization and crystal detection, X-ray diffraction data can be collected 24 days after the start of target gene cloning. Furthermore, improved strategies for serial synchrotron diffraction data collection directly from crystals within living cells abolish the need to purify the recombinant protein or the associated microcrystals.
LB  - PUB:(DE-HGF)16
C6  - pmid:38402242
UR  - <Go to ISI:>//WOS:001177163800012
DO  - DOI:10.1038/s41467-024-45985-7
UR  - https://bib-pubdb1.desy.de/record/605882
ER  -