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A generic protocol for protein crystal dehydration using the HC1b humidity controller

DOI: 10.1107/S2059798316003065 DOI Help

Authors: Carina Lobley (Diamond Light Source) , James Sandy (Diamond Light Source) , Juan Sanchez-weatherby (Diamond Light Source) , Marco Mazzorana (Diamond Light Source) , Tobias Krojer (Oxford University SGC) , Radoslaw Nowak (University of Oxford) , Thomas Sorensen (Diamond Light Source)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Acta Crystallographica Section D Structural Biology , VOL 72 , PAGES 629 - 640

State: Published (Approved)
Published: May 2016

Abstract: Dehydration may change the crystal lattice and affect the mosaicity, resolution and quality of X-ray diffraction data. A dehydrating environment can be generated around a crystal in several ways with various degrees of precision and complexity. This study uses a high-precision crystal humidifier/dehumidifier to provide an airstream of known relative humidity in which the crystals are mounted: a precise yet hassle-free approach to altering crystal hydration. A protocol is introduced to assess the impact of crystal dehydration systematically applied to nine experimental crystal systems. In one case, that of glucose isomerase, dehydration triggering a change of space group from I222 to P21212 was observed. This observation is supported by an extended study of the behaviour of the glucose isomerase crystal structure during crystal dehydration.

Journal Keywords: macromolecular crystallization; crystal dehydration; humidity control; cryoprotection; generic protocol; room temperature.

Subject Areas: Technique Development, Biology and Bio-materials


Instruments: I02-Macromolecular Crystallography