Automation and Experience of Controlled Crystal Dehydration: Results from the European Synchrotron HC1 Collaboration

被引:32
作者
Bowler, Matthew W. [1 ,2 ]
Mueller, Uwe [3 ]
Weiss, Manfred S. [3 ]
Sanchez-Weatherby, Juan [4 ]
Sorensen, Thomas L-M. [4 ]
Thunnissen, Marjolein M. G. M. [5 ,6 ]
Ursby, Thomas [5 ]
Gobbo, Alexandre [1 ,2 ]
Russi, Silvia [1 ,2 ]
Bowler, Michael G. [7 ]
Brockhauser, Sandor [1 ,2 ]
Svensson, Olof [8 ]
Cipriani, Florent [1 ,2 ]
机构
[1] European Mol Biol Lab, Grenoble Outstn, F-38042 Grenoble, France
[2] Univ Grenoble Alpes, EMBL, CNRS, Unit Virus Host Cell Interact, F-38042 Grenoble, France
[3] F ISFM, Helmholtz Zentrum Berlin, Macromol Crystallog HZB MX, D-12489 Berlin, Germany
[4] Diamond Light Source Ltd, Didcot OX11 0DE, Oxon, England
[5] Lund Univ, Max Lab 4, SE-22100 Lund, Sweden
[6] Lund Univ, Ctr Mol Prot Sci, Dept Biochem & Struct Biol, SE-22100 Lund, Sweden
[7] Univ Oxford, Dept Phys, Oxford OX1 3RH, England
[8] European Synchrotron Radiat Facil, Struct Biol Grp, F-38043 Grenoble, France
关键词
MACROMOLECULAR CRYSTALLOGRAPHY; PROTEIN CRYSTALS; RADIATION-DAMAGE; RELATIVE-HUMIDITY; ROOM-TEMPERATURE; DATA-COLLECTION; IMPROVING DIFFRACTION; RNA-POLYMERASE; COMPLEX; BEAMLINE;
D O I
10.1021/cg500890r
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Controlled dehydration of macromolecular crystals can lead to significant improvements in crystalline order, which often manifests itself in higher diffraction quality. Devices that can accurately control the humidity surrounding crystals on a beamline have led to this technique being increasingly adopted as experiments become easier and more reproducible. However, these experiments are often carried out by trial and error, and in order to facilitate and streamline them four European synchrotrons have established a collaboration around the HC1b dehydration device. The MAX IV Laboratory, Diamond Light Source, BESSY II, and the EMBL Grenoble Outstation/ESRF have pooled information gathered from user experiments, and on the use of the device, to propose a set of guidelines for these experiments. Here, we present the status and automation of the installations, advice on how best to perform experiments using the device, and an analysis of successful experiments that begins to show some trends in the type of protocols required by some systems. The dehydration methods shown are applicable to any device that allows control of the relative humidity of the air surrounding a macromolecular crystal.
引用
收藏
页码:1043 / 1054
页数:12
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