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Structure Solution of the Fluorescent Protein Cerulean Using MeshAndCollect

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Hutin, Stephanie | Santoni, Gianluca | Zander, Ulrich | Foos, Nicolas | Aumonier, Sylvain | Gotthard, Guillaume | Royant, Antoine | Mueller-Dieckmann, Christoph | Leonard, Gordon

Edité par CCSD ; JoVE -

International audience. X-ray crystallography is the major technique used to obtain high resolution information concerning the 3-dimensional structures of biological macromolecules. Until recently, a major requirement has been the availability of relatively large, well diffracting crystals, which are often challenging to obtain. However, the advent of serial crystallography and a renaissance in multi-crystal data collection methods has meant that the availability of large crystals need no longer be a limiting factor. Here, we illustrate the use of the automated MeshAndCollect protocol, which first identifies the positions of many small crystals mounted on the same sample holder and then directs the collection from the crystals of a series of partial diffraction data sets for subsequent merging and use in structure determination. MeshAndCollect can be applied to any type of micro-crystals, even if weakly diffracting. As an example, we present here the use of the technique to solve the crystal structure of the Cyan Fluorescent Protein (CFP) Cerulean.

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