Journal Article

PUBDB-2020-04297

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Rapid screening of in cellulo grown protein crystals via a small-angle X-ray scattering/X-ray powder diffraction synergistic approach

Lahey-Rudolph, J. M.Extern*EMBL*DESY* ; Schönherr, R.Extern*DESY* ; Jeffries, C. M.Extern*EMBL* ; Blanchet, C. E.Extern*EMBL* ; Boger, J. ; Ferreira Ramos, A. S.Extern* ; Riekehr, W. M.Extern*EMBL* ; Triandafillidis, D.-P. ; Valmas, A. ; Margiolaki, I. ; Svergun, D.EMBL* ; Redecke, L. (Corresponding author)EMBL*DESY*

2020
Wiley-Blackwell [S.l.]

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Please use a persistent id in citations: doi:10.1107/S1600576720010687  doi:10.3204/PUBDB-2020-04297

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.

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Contributing Institute(s):

1. EMBL-User (EMBL-User)
2. EMBL (EMBL)
3. CFEL-Coherent X-Ray Imaging (FS-CFEL-1)
4. DOOR-User (DOOR ; HAS-User)
5. Universität zu Lübeck (U Lübeck)

Research Program(s):

1. 633 - Life Sciences – Building Blocks of Life: Structure and Function (POF4-633) (POF4-633)
2. 6G3 - PETRA III (POF3-622) (POF3-622)

Experiment(s):

1. PETRA Beamline P12 (PETRA III)

Appears in the scientific report 2020

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Database coverage:
; ; ; Clarivate Analytics Master Journal List ; Current Contents - Physical, Chemical and Earth Sciences ; DEAL Wiley ; Ebsco Academic Search ; Essential Science Indicators ; IF < 5 ; JCR ; NCBI Molecular Biology Database ; Nationallizenz ; PubMed Central ; SCOPUS ; Science Citation Index ; Science Citation Index Expanded ; Web of Science Core Collection

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