Monitoring the Production of High Diffraction-Quality Crystals of Two Enzymes in Real Time Using In Situ Dynamic Light Scattering

Crystals

Claude Sauter¹  Bernard Lorber¹  Kévin Rollet¹  Raphaël de Wijn¹  Philippe Bénas¹  Eric Girard²  Sylvain Engilberge²  Mario Mörl³  Heike Betat³  Oliver Hennig³  AlastairG. McEwen⁴  Olivier Maury⁵  François Riobé⁵ 

[1] Institut de Biologie Moléculaire et Cellulaire, CNRS, UPR 9002, Architecture et Réactivité de l’ARN, Université de Strasbourg, F-67084 Strasbourg, France;Institut de Biologie Structurale, CNRS, CEA, Université Grenoble Alpes, F-38044 Grenoble, France;Institute for Biochemistry, Leipzig University, D-04103 Leipzig, German;Plateforme de Biologie et Génomique Structurale, Institut de Génétique et de Biologie Moléculaire et Cellulaire, Université de Strasbourg, CNRS UMR 7104, INSERM U 1258, F-67404 Illkirch, France;Univ Lyon, ENS Lyon, CNRS-UMR 5182, Université Lyon 1, F-69342 Lyon, France;

关键词: enzyme;    crystallization;    dynamic light scattering;    nucleation;    nucleant;    tb-xo4 crystallophore;    microcrystals;    nanocrystals;    x-ray diffraction;    xtalcontroller;   

DOI  :  10.3390/cryst10020065

来源: DOAJ

【 摘 要 】

The reproducible preparation of well-diffracting crystals is a prerequisite for every structural study based on crystallography. An instrument called XtalController has recently been designed that allows the monitoring of crystallization assays using dynamic light scattering and microscopy, and integrates piezo pumps to alter the composition of the mother liquor during the experiment. We have applied this technology to study the crystallization of two enzymes, the CCA-adding enzyme of the psychrophilic bacterium Planococcus halocryophilus, and the lysozyme from hen egg white in the presence of a synthetic chemical nucleant. We were able to (i) detect early nucleation events and (ii) drive the crystallization system (through cycles of dissolution/crystallization) toward growth conditions yielding crystals with excellent diffraction properties. This technology opens a way to the rational production of samples for crystallography, ranging from nanocrystals for electron diffraction, microcrystals for serial or conventional X-ray diffraction, to larger crystals for neutron diffraction.

【 授权许可】

Unknown