Automated analysis of vapor diffusion crystallization drops with an X-ray beam.
Crystallogenesis, usually based on the vapor diffusion method, is currently considered one of the most difficult steps in macromolecular X-ray crystallography. Due to the increasing number of crystallization assays performed by protein crystallographers, several automated analysis methods are under development. Most of these methods are based on microscope images and shape recognition. We propose an alternative method of identifying protein crystals: by directly exposing the crystallization drops to an X-ray beam. The resulting diffraction provides far more information than classical microscope images. Not only is the presence of diffracting crystals revealed, but also a first estimation of the space group, cell parameters, and mosaicity is obtained. In certain cases, it is also possible to collect enough data to verify the presence of a specific substrate or a heavy atom. All these steps are performed without the sometimes tedious necessity of removing crystals from their crystallization drop.
Jacquamet
Lilian
L
IBS J.-P. Ebel CEA-CNRS-UJF, 41 rue Jules Horowitz, 38027 Grenoble Cedex 1, France.
Ohana
Jeremy
J
Joly
Jacques
J
Borel
Franck
F
Pirocchi
Michel
M
Charrault
Philippe
P
Bertoni
Alain
A
Israel-Gouy
Pascale
P
Carpentier
Philippe
P
Kozielski
Frank
F
Blot
Delphine
D
Ferrer
Jean-Luc
JL
D003460:Crystallization; D018360:Crystallography, X-Ray / Q000295:instrumentation / Q000379:methods*; D003625:Data Collection; D004867:Equipment Design; D011506:Proteins / Q000737:chemistry*; D012371:Robotics
D011506:Proteins