title: In cellulo serial crystallography of alcohol oxidase crystals inside yeast cells.
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.
authors.affiliation: Hamburg Unit c/o DESY, European Molecular Biology Laboratory (EMBL), Notkestrasse 85, 22607 Hamburg, Germany; Structural and Computational Biology Unit, European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, 69117 Heidelberg, Germany.
authors.forename: Arjen J
authors.initials: AJ
authors.lastname: Jakobi
authors.affiliation: Hamburg Unit c/o DESY, European Molecular Biology Laboratory (EMBL) , Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Passon
authors.affiliation: Molecular Cell Biology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen , 9747 AG Groningen, The Netherlands.
authors.forename: Kèvin
authors.initials: K
authors.lastname: Knoops
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen Synchrotron DESY , Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Francesco
authors.initials: F
authors.lastname: Stellato
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen Synchrotron DESY , Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Mengning
authors.initials: M
authors.lastname: Liang
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen Synchrotron DESY , Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: White
authors.affiliation: Hamburg Unit c/o DESY, European Molecular Biology Laboratory (EMBL), Notkestrasse 85, 22607 Hamburg, Germany; Center for Free-Electron Laser Science, Deutsches Elektronen Synchrotron DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Seine
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory , Menlo Park, California, USA.
authors.forename: Marc
authors.initials: M
authors.lastname: Messerschmidt
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen Synchrotron DESY, Notkestrasse 85, 22607 Hamburg, Germany; Department of Physics, University of Hamburg, Luruper Chaussee 149, 22607 Hamburg, Germany; Center for Ultrafast Imaging, Luruper Chaussee 149, 22607 Hamburg, Germany.
authors.forename: Henry N
authors.initials: HN
authors.lastname: Chapman
authors.affiliation: Hamburg Unit c/o DESY, European Molecular Biology Laboratory (EMBL), Notkestrasse 85, 22607 Hamburg, Germany; University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Wilmanns
keywords: X-ray crystallography; femtosecond studies; free-electron laser; nanocrystals; protein structure