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Fast high-pressure freezing of protein crystals in their mother liquor

DOI: 10.1107/S1744309112009670 DOI Help
PMID: 22505429 PMID Help

Authors: Anja Burkhardt (Deutsches Elektronen-Synchrotron DESY, Germany) , Martin Warmer (Leibniz Institute for Experimental Virology, Germany) , Saravanan Panneerselvam (Deutsches Elektronen-Synchrotron DESY, Germany) , Armin Wagner (Diamond Light Source) , Athina Zouni (Technical University Berlin, Germany) , Carina Gloeckner (Technical University Berlin, Germany) , Rudolph Reimer (Leibniz Institute for Experimental Virology, Germany) , Heinrich Hohenberg (Leibniz Institute for Experimental Virology, Germany) , Alke Meents (Deutsches Elektronen-Synchrotron DESY, Germany)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Acta Crystallographica Section F Structural Biology And Crystallization Communications , VOL 68 (4) , PAGES 495-500

State: Published (Approved)
Published: April 2012

Abstract: High-pressure freezing (HPF) is a method which allows sample vitrification without cryoprotectants. In the present work, protein crystals were cooled to cryogenic temperatures at a pressure of 210 MPa. In contrast to other HPF methods published to date in the field of cryocrystallography, this protocol involves rapid sample cooling using a standard HPF device. The fast cooling rates allow HPF of protein crystals directly in their mother liquor without the need for cryoprotectants or external reagents. HPF was first attempted with hen egg-white lysozyme and cubic insulin crystals, yielding good to excellent diffraction quality. Non-cryoprotected crystals of the membrane protein photosystem II have been successfully cryocooled for the first time. This indicates that the presented HPF method is well suited to the vitrification of challenging systems with large unit cells and weak crystal contacts.

Journal Keywords: Chickens; Crystallography; X-Ray; Freezing; Insulin; Muramidase; Pressure; Time Factors; Cooling; Crystals; Diffraction; Equipment; Freezing; Lysozyme; Pressure Range Mega Pa 10-100; Vitrification

Subject Areas: Technique Development, Biology and Bio-materials

Instruments: I04-1-Macromolecular Crystallography (fixed wavelength) , I24-Microfocus Macromolecular Crystallography