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Birth and Growth Kinetics of Brome Mosaïc Virus Microcrystals

Marina Casselyn ^*, Annette Tardieu , Hervé Delacroix ^* and Stéphanie Finet 

^* BioInformatique Structurale, Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique, Gif-sur-Yvette Cedex, France; and Laboratoire de Minéralogie et Cristallographie de Paris, Centre National de la Recherche Scientifique, Université Paris VI Pierre et Marie Curie, Paris, France; and European Synchrotron Radiation Facility, BP 220, Grenoble, France

Correspondence: Address reprint requests to Dr. Stéphanie Finet, ESRF, BP 220, 38043 Grenoble Cedex, France. Tel.: 33-476-88-2682; Fax: 33-476-88-2325; E-mail: finet{at}esrf.fr.

The early steps of crystal nucleation and growth in Brome Mosaïc virus and polyethylene glycol mixtures were analyzed using time-resolved x-ray scattering (at the European Synchrotron Radiation Facility, Grenoble, France). The system was chosen as a crystallization model since the phase diagram of the macromolecule/polymer mixture was known to present, at high polymer concentration, a solid, precipitated phase made of the synchronized formation of a large number of microcrystals. The precipitation and crystallization of the samples was induced by the controlled mixing of virus and polymer using a stopped-flow device. Appearance and growth of Bragg diffraction peaks were used to follow the crystal nucleation and growth as a function of time, virus and polymer concentration, and polymer size. In all samples, the crystallization starts after a few seconds and proceeds for 1–20 min until there is almost no virus left in the solution. The crystalline system was found to be face-centered cubic, with a unit cell size of 391 Å. The data analysis allowed us to show the presence of viruses in only two states, in solution or in crystals, revealing that the formation of periodic order proceeds without any detectable intermediate amorphous state.