Journal
IUCRJ
Volume 3, Issue -, Pages 88-95Publisher
INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S2052252515022927
Keywords
protein structure; X-ray crystallography; femtosecond studies; nanocrystals; free-electron laser
Funding
- NSF [1231306]
- Marie Sklodowska-Curie IEF grant [FP7-IEF-330150, PIEF-GA-2012-331285]
- EMBL Interdisciplinary Post-doctoral (EIPOD) fellowship under Marie Sklodowska-Curie COFUND Actions [PCOFUND-GA-2008-229597]
- Hamburg Center for Ultrafast Imaging - Structure, Dynamics and Control of Matter at the Atomic Scale centre of excellence of the Deutsche Forschungsgemeinschaft
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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.
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