Structure Solution of the Fluorescent Protein Cerulean Using MeshAndCollect.


Journal

Journal of visualized experiments : JoVE
ISSN: 1940-087X
Titre abrégé: J Vis Exp
Pays: United States
ID NLM: 101313252

Informations de publication

Date de publication:
19 03 2019
Historique:
entrez: 9 4 2019
pubmed: 9 4 2019
medline: 31 1 2020
Statut: epublish

Résumé

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.

Identifiants

pubmed: 30958477
doi: 10.3791/58594
doi:

Substances chimiques

Cyan Fluorescent Protein 0
Green Fluorescent Proteins 147336-22-9

Types de publication

Journal Article Video-Audio Media

Langues

eng

Sous-ensembles de citation

IM

Auteurs

Stephanie Hutin (S)

European Synchrotron Radiation Facility, Structural Biology Group; stephanie.hutin@esrf.fr.

Gianluca Santoni (G)

European Synchrotron Radiation Facility, Structural Biology Group.

Ulrich Zander (U)

European Molecular Biology Laboratory.

Nicolas Foos (N)

European Synchrotron Radiation Facility, Structural Biology Group.

Sylvain Aumonier (S)

European Synchrotron Radiation Facility, Structural Biology Group.

Guillaume Gotthard (G)

European Synchrotron Radiation Facility, Structural Biology Group.

Antoine Royant (A)

European Synchrotron Radiation Facility, Structural Biology Group; Univ. Grenoble Alpes, CNRS, CEA, IBS (Institut de Biologie Structurale).

Christoph Mueller-Dieckmann (C)

European Synchrotron Radiation Facility, Structural Biology Group.

Gordon Leonard (G)

European Synchrotron Radiation Facility, Structural Biology Group.

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Classifications MeSH