8C5K

HEX-1 (in cellulo, in situ) crystallized and diffracted in High Five cells. Growth and SX data collection at 296 K on CrystalDirect plates

Summary for 8C5K

• Entry DOI: 10.2210/pdb8c5k/pdb
• Related: 7ASX 8C51 8C53
• Descriptor: Woronin body major protein (2 entities in total)
• Functional Keywords: woronin body, self-assembly, hex-1, in vivo, structural protein
• Biological source: Neurospora crassa OR74A
• Total number of polymer chains: 1
• Total formula weight: 19150.66

Authors

Lahey-Rudolph, J.M.,Schoenherr, R.,Boger, J.,Harms, M.,Kaiser, J.,Nachtschatt, S.,Wobbe, M.,Duden, R.,Koenig, P.,Bourenkov, G.,Schneider, T.,Redecke, L. (deposition date: 2023-01-09, release date: 2024-01-17, Last modification date: 2024-03-06)

Primary citation

Schonherr, R.,Boger, J.,Lahey-Rudolph, J.M.,Harms, M.,Kaiser, J.,Nachtschatt, S.,Wobbe, M.,Duden, R.,Konig, P.,Bourenkov, G.,Schneider, T.R.,Redecke, L.
A streamlined approach to structure elucidation using in cellulo crystallized recombinant proteins, InCellCryst.
Nat Commun, 15:1709-1709, 2024

PubMed Abstract: 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.

PubMed: 38402242
DOI: 10.1038/s41467-024-45985-7

Experimental method

X-RAY DIFFRACTION (2.16 Å)