Monitoring the Production of High Diffraction-Quality Crystals of Two Enzymes in Real Time Using In Situ Dynamic Light Scattering
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 inte...
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doaj-b882c0ad037445599b4fa616cf9fd8402020-11-25T01:45:51ZengMDPI AGCrystals2073-43522020-01-011026510.3390/cryst10020065cryst10020065Monitoring the Production of High Diffraction-Quality Crystals of Two Enzymes in Real Time Using In Situ Dynamic Light ScatteringRaphaël de Wijn0Kévin Rollet1Sylvain Engilberge2Alastair G. McEwen3Oliver Hennig4Heike Betat5Mario Mörl6François Riobé7Olivier Maury8Eric Girard9Philippe Bénas10Bernard Lorber11Claude Sauter12Institut de Biologie Moléculaire et Cellulaire, CNRS, UPR 9002, Architecture et Réactivité de l’ARN, Université de Strasbourg, F-67084 Strasbourg, FranceInstitut de Biologie Moléculaire et Cellulaire, CNRS, UPR 9002, Architecture et Réactivité de l’ARN, Université de Strasbourg, F-67084 Strasbourg, FranceInstitut de Biologie Structurale, CNRS, CEA, Université Grenoble Alpes, F-38044 Grenoble, FrancePlateforme 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, FranceInstitute for Biochemistry, Leipzig University, D-04103 Leipzig, GermanInstitute for Biochemistry, Leipzig University, D-04103 Leipzig, GermanInstitute for Biochemistry, Leipzig University, D-04103 Leipzig, GermanUniv Lyon, ENS Lyon, CNRS-UMR 5182, Université Lyon 1, F-69342 Lyon, FranceUniv Lyon, ENS Lyon, CNRS-UMR 5182, Université Lyon 1, F-69342 Lyon, FranceInstitut de Biologie Structurale, CNRS, CEA, Université Grenoble Alpes, F-38044 Grenoble, FranceInstitut de Biologie Moléculaire et Cellulaire, CNRS, UPR 9002, Architecture et Réactivité de l’ARN, Université de Strasbourg, F-67084 Strasbourg, FranceInstitut de Biologie Moléculaire et Cellulaire, CNRS, UPR 9002, Architecture et Réactivité de l’ARN, Université de Strasbourg, F-67084 Strasbourg, FranceInstitut de Biologie Moléculaire et Cellulaire, CNRS, UPR 9002, Architecture et Réactivité de l’ARN, Université de Strasbourg, F-67084 Strasbourg, FranceThe 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 <i>Planococcus halocryophilus,</i> 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.https://www.mdpi.com/2073-4352/10/2/65enzymecrystallizationdynamic light scatteringnucleationnucleanttb-xo4 crystallophoremicrocrystalsnanocrystalsx-ray diffractionxtalcontroller |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Raphaël de Wijn Kévin Rollet Sylvain Engilberge Alastair G. McEwen Oliver Hennig Heike Betat Mario Mörl François Riobé Olivier Maury Eric Girard Philippe Bénas Bernard Lorber Claude Sauter |
spellingShingle |
Raphaël de Wijn Kévin Rollet Sylvain Engilberge Alastair G. McEwen Oliver Hennig Heike Betat Mario Mörl François Riobé Olivier Maury Eric Girard Philippe Bénas Bernard Lorber Claude Sauter Monitoring the Production of High Diffraction-Quality Crystals of Two Enzymes in Real Time Using In Situ Dynamic Light Scattering Crystals enzyme crystallization dynamic light scattering nucleation nucleant tb-xo4 crystallophore microcrystals nanocrystals x-ray diffraction xtalcontroller |
author_facet |
Raphaël de Wijn Kévin Rollet Sylvain Engilberge Alastair G. McEwen Oliver Hennig Heike Betat Mario Mörl François Riobé Olivier Maury Eric Girard Philippe Bénas Bernard Lorber Claude Sauter |
author_sort |
Raphaël de Wijn |
title |
Monitoring the Production of High Diffraction-Quality Crystals of Two Enzymes in Real Time Using In Situ Dynamic Light Scattering |
title_short |
Monitoring the Production of High Diffraction-Quality Crystals of Two Enzymes in Real Time Using In Situ Dynamic Light Scattering |
title_full |
Monitoring the Production of High Diffraction-Quality Crystals of Two Enzymes in Real Time Using In Situ Dynamic Light Scattering |
title_fullStr |
Monitoring the Production of High Diffraction-Quality Crystals of Two Enzymes in Real Time Using In Situ Dynamic Light Scattering |
title_full_unstemmed |
Monitoring the Production of High Diffraction-Quality Crystals of Two Enzymes in Real Time Using In Situ Dynamic Light Scattering |
title_sort |
monitoring the production of high diffraction-quality crystals of two enzymes in real time using in situ dynamic light scattering |
publisher |
MDPI AG |
series |
Crystals |
issn |
2073-4352 |
publishDate |
2020-01-01 |
description |
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 <i>Planococcus halocryophilus,</i> 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. |
topic |
enzyme crystallization dynamic light scattering nucleation nucleant tb-xo4 crystallophore microcrystals nanocrystals x-ray diffraction xtalcontroller |
url |
https://www.mdpi.com/2073-4352/10/2/65 |
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