Published in
International Union of Crystallography, IUCrJ, 2(3), p. 88-95, 2016
DOI: 10.1107/s2052252515022927
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In celluloserial crystallography of alcohol oxidase crystals inside yeast cells
,
Francesco Stellato,
Mengning Liang,
Thomas A. White ,
Thomas Seine,
Marc Messerschmidt,
Henry N. Chapman ,
Matthias Wilmanns
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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 yeastHansenula polymorphais reported and characterized. The observations are supported by synchrotron radiation-based powder diffraction data and electron microscopy. Based on these findings, the concept ofin celluloserial crystallography on protein targets imported into yeast peroxisomes without the need for protein purification as a requirement for subsequent crystallization is outlined.