De novo protein crystal structure determination from X-ray free-electron laser data

Thomas R M Barends, Lutz Foucar, Sabine Botha, R. Bruce Doak, Robert L. Shoeman, Karol Nass, Jason E. Koglin, Garth J. Williams, Sébastien Boutet, Marc Messerschmidt, Ilme Schlichting

Research output: Contribution to journalArticle

186 Citations (Scopus)

Abstract

The determination of protein crystal structures is hampered by the need for macroscopic crystals. X-ray free-electron lasers (FELs) provide extremely intense pulses of femtosecond duration, which allow data collection from nanometre-to micrometre-sized crystals in a 'diffraction-before-destruction' approach. So far, all protein structure determinations carried out using FELs have been based on previous knowledge of related, known structures. Here we show that X-ray FEL data can be used for de novo protein structure determination, that is, without previous knowledge about the structure. Using the emerging technique of serial femtosecond crystallography, we performed single-wavelength anomalous scattering measurements on microcrystals of the well-established model system lysozyme, in complex with a lanthanide compound. Using Monte-Carlo integration, we obtained high-quality diffraction intensities from which experimental phases could be determined, resulting in an experimental electron density map good enough for automated building of the protein structure. This demonstrates the feasibility of determining novel protein structures using FELs. We anticipate that serial femtosecond crystallography will become an important tool for the structure determination of proteins that are difficult to crystallize, such as membrane proteins.

Original languageEnglish (US)
Pages (from-to)244-247
Number of pages4
JournalNature
Volume505
Issue number7482
DOIs
StatePublished - Jan 1 2014
Externally publishedYes

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Lasers
X-Rays
Electrons
Crystallography
Proteins
Lanthanoid Series Elements
Muramidase
Membrane Proteins

ASJC Scopus subject areas

  • General

Cite this

Barends, T. R. M., Foucar, L., Botha, S., Doak, R. B., Shoeman, R. L., Nass, K., ... Schlichting, I. (2014). De novo protein crystal structure determination from X-ray free-electron laser data. Nature, 505(7482), 244-247. https://doi.org/10.1038/nature12773

De novo protein crystal structure determination from X-ray free-electron laser data. / Barends, Thomas R M; Foucar, Lutz; Botha, Sabine; Doak, R. Bruce; Shoeman, Robert L.; Nass, Karol; Koglin, Jason E.; Williams, Garth J.; Boutet, Sébastien; Messerschmidt, Marc; Schlichting, Ilme.

In: Nature, Vol. 505, No. 7482, 01.01.2014, p. 244-247.

Research output: Contribution to journalArticle

Barends, TRM, Foucar, L, Botha, S, Doak, RB, Shoeman, RL, Nass, K, Koglin, JE, Williams, GJ, Boutet, S, Messerschmidt, M & Schlichting, I 2014, 'De novo protein crystal structure determination from X-ray free-electron laser data', Nature, vol. 505, no. 7482, pp. 244-247. https://doi.org/10.1038/nature12773
Barends TRM, Foucar L, Botha S, Doak RB, Shoeman RL, Nass K et al. De novo protein crystal structure determination from X-ray free-electron laser data. Nature. 2014 Jan 1;505(7482):244-247. https://doi.org/10.1038/nature12773
Barends, Thomas R M ; Foucar, Lutz ; Botha, Sabine ; Doak, R. Bruce ; Shoeman, Robert L. ; Nass, Karol ; Koglin, Jason E. ; Williams, Garth J. ; Boutet, Sébastien ; Messerschmidt, Marc ; Schlichting, Ilme. / De novo protein crystal structure determination from X-ray free-electron laser data. In: Nature. 2014 ; Vol. 505, No. 7482. pp. 244-247.
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