Protein crystal structure obtained at 2.9 Å resolution from injecting bacterial cells into an X-ray free-electron laser beam

Michael R. Sawaya, Duilio Cascio, Mari Gingery, Jose Rodriguez, Lukasz Goldschmidt, Jacques Philippe Colletier, Marc Messerschmidt, Sébastien Boutet, Jason E. Koglin, Garth J. Williams, Aaron S. Brewster, Karol Nass, Johan Hattne, Sabine Botha, R. Bruce Doak, Robert L. Shoeman, Daniel P. DePonte, Hyun Woo Park, Brian A. Federici, Nicholas K. SauterIlme Schlichting, David S. Eisenberg

Research output: Contribution to journalArticle

67 Scopus citations

Abstract

It has long been known that toxins produced by Bacillus thuringiensis (Bt) are stored in the bacterial cells in crystalline form. Here we describe the structure determination of the Cry3A toxin found naturally crystallized within Bt cells. When whole Bt cells were streamed into an X-ray free-electron laser beam we found that scattering from other cell components did not obscure diffraction fromthe crystals. The resolution limits of the best diffraction images collected from cells were the same as from isolated crystals. The integrity of the cells at the moment of diffraction is unclear; however, given the short time (?5 μs) between exiting the injector to intersecting with the X-ray beam, our result is a 2.9-Å-resolution structure of a crystalline protein as it exists in a living cell. The study suggests that authentic in vivo diffraction studies can produce atomic-level structural information.

Original languageEnglish (US)
Pages (from-to)12769-12774
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume111
Issue number35
DOIs
StatePublished - Sep 2 2014
Externally publishedYes

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Keywords

  • Cry3A insecticidal toxin
  • Serial femtosecond crystallography
  • XFEL

ASJC Scopus subject areas

  • General

Cite this

Sawaya, M. R., Cascio, D., Gingery, M., Rodriguez, J., Goldschmidt, L., Colletier, J. P., Messerschmidt, M., Boutet, S., Koglin, J. E., Williams, G. J., Brewster, A. S., Nass, K., Hattne, J., Botha, S., Doak, R. B., Shoeman, R. L., DePonte, D. P., Park, H. W., Federici, B. A., ... Eisenberg, D. S. (2014). Protein crystal structure obtained at 2.9 Å resolution from injecting bacterial cells into an X-ray free-electron laser beam. Proceedings of the National Academy of Sciences of the United States of America, 111(35), 12769-12774. https://doi.org/10.1073/pnas.1413456111