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In cellulo serial crystallography of alcohol oxidase crystals inside yeast cells

By Arjen J. Jakobi, Daniel M. Passon, Kèvin Knoops, Francesco Stellato, Mengning Liang, Thomas A. White, Thomas Seine, Marc Messerschmidt1, Henry Chapman2, Matthias Wilmanns

1. Arizona State University 2. Center for Free-Electron Laser Science

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Category

BioXFEL Publications

Published on

Type

journal-article

Author

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

Citation

Jakobi, A.J. et al., 2016. In cellulo serial crystallography of alcohol oxidase crystals inside yeast cells . IUCrJ, 3(2). Available at: http://dx.doi.org/10.1107/s2052252515022927.

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.

DOI

http://dx.doi.org/10.1107/s2052252515022927

Funding

NSF-STC Biology with X-ray Lasers (NSF-1231306)