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  A standardized technique for high-pressure cooling of protein crystals

Quirnheim Pais, D., Rathmann, B., Koepke, J., Tomova, C., Wurzinger, P., & Thielmann, Y. (2017). A standardized technique for high-pressure cooling of protein crystals. Acta Crystallographica. Section D: Biological Crystallography (Copenhagen), 73(12), 997-1006. doi:10.1107/S2059798317016357.

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 Creators:
Quirnheim Pais, David1, Author           
Rathmann, Barbara1, Author           
Koepke, Jürgen1, Author           
Tomova, Cveta2, Author
Wurzinger, Paul2, Author
Thielmann, Yvonne1, Author           
Affiliations:
1Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society, ou_2068290              
2Leica Microsystems Vienna, Hernalser Hauptstrasse 219, 1170 Vienna, Austria, ou_persistent22              

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Free keywords: high-pressure cooling; protein crystals; polyimide capillary; buffer composition
 Abstract: Cryogenic temperatures slow down secondary radiation damage during data collection from macromolecular crystals. In 1973, cooling at high pressure was identified as a method for cryopreserving crystals in their mother liquor [Thomanek et al. (1973). Acta Cryst. A29, 263–265]. Results from different groups studying different crystal systems indicated that the approach had merit, although difficulties in making the process work have limited its widespread use. Therefore, a simplified and reliable technique has been developed termed high-pressure cooling (HPC). An essential requirement for HPC is to protect crystals in capillaries. These capillaries form part of new sample holders with SPINE standard dimensions. Crystals are harvested with the capillary, cooled at high pressure (220 MPa) and stored in a cryovial. This system also allows the usage of the standard automation at the synchrotron. Crystals of hen egg-white lysozyme and concanavalin A have been successfully cryopreserved and yielded data sets to resolutions of 1.45 and 1.35 Å, respectively. Extensive work has been performed to define the useful working range of HPC in capillaries with 250 µm inner diameter. Three different 96-well crystallization screens that are most frequently used in our crystallization facility were chosen to study the formation of amorphous ice in this cooling setup. More than 89% of the screening solutions were directly suitable for HPC. This achievement represents a drastic improvement for crystals that suffered from cryoprotection or were not previously eligible for cryoprotection.

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Language(s): eng - English
 Dates: 2017-06-162017-11-142017-11-222017-12
 Publication Status: Published in print
 Pages: 10
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1107/S2059798317016357
 Degree: -

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Title: Acta Crystallographica. Section D: Biological Crystallography (Copenhagen)
  Other : Acta Crystallogr D
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: [Copenhagen, Denmark : Published for the International Union of Crystallography by Munksgaard]
Pages: - Volume / Issue: 73 (12) Sequence Number: - Start / End Page: 997 - 1006 Identifier: ISSN: 0907-4449
CoNE: https://pure.mpg.de/cone/journals/resource/954925562619