Counterdiffusion protein crystallisation in microgravity and its observation with PromISS (protein microscope for the international space station) |
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Authors: | Ingrid Zegers Luigi Carotenuto Christine Evrard JuanMa Garcia-Ruiz Philippe De Gieter Luis Gonzales-Ramires Eric Istasse Jean-Claude Legros Joseph Martial Christophe Minetti Fermin Otalora Patrick Queeckers Cedric Schockaert Cecile VandeWeerdt Ronnie Willaert Lode Wyns Catherine Yourassowsky Frank Dubois |
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Affiliation: | 1. Department Ultrastructure, VUB. Pleinlaan 2, 1050, Brussels, Belgium 2. MARS Center, Via Emanuele Gianturco 31, 80146, Napoli, Italy 3. UCL-CSTR, Place Louis Pasteur 1, 1348, Lovain-la-Neuve, Belgium 4. Laboratorio de Estudios Cristalográficos, Edificio BIC Granada, Avenida de la Innovación, 1, E-18100, Armilla, Granada, Spain 5. Microgravity Research centre, ULB, Avenue F.D. Roosevelt 50, 1050, Bruxelles, Belgium 6. Labo de biologie moléculaire et de génie génétique, Bat. B6, Allée de la Chimie, 3, 4000, Liège, Belgium
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Abstract: | The crystallisation by counterdiffusion is a very efficient technique for obtaining high-quality protein crystals. A prerequisite for the use of counterdiffusion techniques is that mass transport must be controlled by diffusion alone. Sedimentation and convection can be avoided by either working in gelled systems, working in systems of small dimensions, or in the absence of gravity. We present the results from experiments performed on the ISS using the Protein Microscope for the International Space Station (PromISS), using digital holography to visualise crystal growth processes. We extensively characterised three model proteins for these experiments (cablys3*lysozyme, triose phosphate isomerase, and parvalbumin) and used these to assess the ISS as an environment for crystallisation by counterdiffusion. The possibility to visualise growth and movement of crystals in different types of experiments (capillary counterdiffusion and batch-type) is important, as movement of crystals is clearly not negligible. |
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