In cellulo serial crystallography of alcohol oxidase crystals inside yeast cells

Arjen J. Jakobi, Daniel M. Passon, Kèvin Knoops, Francesco Stellato, Mengning Liang, Thomas A. White, Thomas Seine, Marc Messerschmidt, Henry N. Chapman, Matthias Wilmanns

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

The possibility of using femtosecond pulses from an X-ray free-electron laser to collect diffraction data from protein crystals formed in their native cellular organelle has been explored. X-ray diffraction of submicrometre-sized alcohol oxidase crystals formed in peroxisomes within cells of genetically modified variants of the methylotrophic yeast Hansenula polymorpha is reported and characterized. The observations are supported by synchrotron radiation-based powder diffraction data and electron microscopy. Based on these findings, the concept of in cellulo serial crystallography on protein targets imported into yeast peroxisomes without the need for protein purification as a requirement for subsequent crystallization is outlined.

Original languageEnglish (US)
Pages (from-to)88-95
Number of pages8
JournalIUCrJ
Volume3
DOIs
StatePublished - Jan 1 2016
Externally publishedYes

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Keywords

  • femtosecond studies
  • free-electron laser
  • nanocrystals
  • protein structure
  • X-ray crystallography

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry
  • Materials Science(all)
  • Condensed Matter Physics

Cite this

Jakobi, A. J., Passon, D. M., Knoops, K., Stellato, F., Liang, M., White, T. A., Seine, T., Messerschmidt, M., Chapman, H. N., & Wilmanns, M. (2016). In cellulo serial crystallography of alcohol oxidase crystals inside yeast cells. IUCrJ, 3, 88-95. https://doi.org/10.1107/S2052252515022927