Rapid screening of in cellulo grown protein crystals via a small-angle X-ray scattering/X-ray powder diffraction synergistic approach

© Janine Mia Lahey-Rudolph et al. 2020.

Bibliographische Detailangaben
Veröffentlicht in:Journal of applied crystallography. - 1998. - 53(2020), Pt 5 vom: 01. Okt., Seite 1169-1180
1. Verfasser: Lahey-Rudolph, Janine Mia (VerfasserIn)
Weitere Verfasser: Schönherr, Robert, Jeffries, Cy M, Blanchet, Clément E, Boger, Juliane, Ferreira Ramos, Ana Sofia, Riekehr, Winnie Maria, Triandafillidis, Dimitris-Panagiotis, Valmas, Alexandros, Margiolaki, Irene, Svergun, Dmitri, Redecke, Lars
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2020
Zugriff auf das übergeordnete Werk:Journal of applied crystallography
Schlagworte:Journal Article X-ray powder diffraction in cellulo crystals protein micro-crystallography small-angle X-ray scattering
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520 |a 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 
650 4 |a Journal Article 
650 4 |a X-ray powder diffraction 
650 4 |a in cellulo crystals 
650 4 |a protein micro-crystallography 
650 4 |a small-angle X-ray scattering 
700 1 |a Schönherr, Robert  |e verfasserin  |4 aut 
700 1 |a Jeffries, Cy M  |e verfasserin  |4 aut 
700 1 |a Blanchet, Clément E  |e verfasserin  |4 aut 
700 1 |a Boger, Juliane  |e verfasserin  |4 aut 
700 1 |a Ferreira Ramos, Ana Sofia  |e verfasserin  |4 aut 
700 1 |a Riekehr, Winnie Maria  |e verfasserin  |4 aut 
700 1 |a Triandafillidis, Dimitris-Panagiotis  |e verfasserin  |4 aut 
700 1 |a Valmas, Alexandros  |e verfasserin  |4 aut 
700 1 |a Margiolaki, Irene  |e verfasserin  |4 aut 
700 1 |a Svergun, Dmitri  |e verfasserin  |4 aut 
700 1 |a Redecke, Lars  |e verfasserin  |4 aut 
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