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Attaining atomic resolution from in situ data collection at room temperature using counter-diffusion-based low-cost microchips

Gavira, Jose A. and Rodriguez-Ruiz, Isaac and Martinez-Rodriguez, Sergio and Basu, Shibom and Teychené, Sébastien and McCarthy, Andrew A. and Mueller-Dieckman, Christoph Attaining atomic resolution from in situ data collection at room temperature using counter-diffusion-based low-cost microchips. (2020) Acta Crystallographica Section D Structural Biology, 76 (8). 751-758. ISSN 2059-7983

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Official URL: https://doi.org/10.1107/S2059798320008475

Abstract

Sample handling and manipulation for cryoprotection currently remain critical factors in X-ray structural determination. While several microchips for macromolecular crystallization have been proposed during the last two decades to partially overcome crystal-manipulation issues, increased background noise originating from the scattering of chip-fabrication materials has so far limited the attainable resolution of diffraction data. Here, the conception and use of low-cost, X-ray-transparent microchips for in situ crystallization and direct data collection, and structure determination at atomic resolution close to 1.0 Å , is presented. The chips are fabricated by a combination of either OSTEMER and Kapton or OSTEMER and Mylar materials for the implementation of counter-diffusion crystallization experiments. Both materials produce a sufficiently low scattering background to permit atomic resolution diffraction data collection at room temperature and the generation of 3D structural models of the tested model proteins lysozyme, thaumatin and glucose isomerase. Although the high symmetry of the three model protein crystals produced almost complete data sets at high resolution, the potential of in-line data merging and scaling of the multiple crystals grown along the microfluidic channels is also presented and discussed.

Item Type:Article
Audience (journal):International peer-reviewed journal
Uncontrolled Keywords:
Institution:French research institutions > Centre National de la Recherche Scientifique - CNRS (FRANCE)
Other partners > European Molecular Biology Laboratory - EMBL (FRANCE)
Other partners > European Synchrotron Radiation Facility - ESRF (FRANCE)
Université de Toulouse > Institut National Polytechnique de Toulouse - Toulouse INP (FRANCE)
Université de Toulouse > Université Toulouse III - Paul Sabatier - UT3 (FRANCE)
Other partners > Universidad de Granada - UGR (SPAIN)
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Deposited On:15 Feb 2021 13:07

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