8CGY

Trypanosoma brucei IMP dehydrogenase (ori) crystallized in High Five cells reveals native ligands ATP, GDP and phosphate. Diffraction data collection at 100 K in cellulo; XDS processing

8CGY の概要

• エントリーDOI: 10.2210/pdb8cgy/pdb
• 分子名称: Inosine-5'-monophosphate dehydrogenase, ADENOSINE-5'-TRIPHOSPHATE, GUANOSINE-5'-DIPHOSPHATE, ... (5 entities in total)
• 機能のキーワード: imp dehydrogenase, purine biosynthetic enzyme, native cofactors, transferase
• 由来する生物種: Trypanosoma brucei
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 113622.02

構造登録者

Boger, J.,Schoenherr, R.,Lahey-Rudolph, J.M.,Harms, M.,Kaiser, J.,Nachtschatt, S.,Wobbe, M.,Duden, R.,Bourenkov, G.,Schneider, T.,Redecke, L. (登録日: 2023-02-06, 公開日: 2024-02-21, 最終更新日: 2024-03-06)

主引用文献

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

実験手法

X-RAY DIFFRACTION (3 Å)