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Diagnostic of precipitant for biomacromolecule crystallization by quasi-elastic light-scattering

Published on May 1, 1990in Journal of Molecular Biology5.067
· DOI :10.1016/S0022-2836(05)80130-5
Vincent Mikol7
Estimated H-index: 7
(CNRS: Centre national de la recherche scientifique),
Ernest Hirsch6
Estimated H-index: 6
,
Richard Giegé54
Estimated H-index: 54
(CNRS: Centre national de la recherche scientifique)
Abstract
The translational diffusion coefficient D 25,w of hen egg-white lysozyme and concanavalin A from the jack bean is measured in various precipitating agent solutions as a function of salt and protein concentration using quasi-elastic light-scattering. With some precipitants, in undersaturated protein solutions, a protein or salt concentration dependence of the diffision coefficient of the scatterers is observed. It can be correlated with the inability of the protein to crystallize in this precipitant once the solution is supersaturated. These variations of D 25,w are interpreted in terms of non-specific interactions and/or aggregation that prevent the protein from making appropriate contacts to form a crystal. With other precipitants known to lead to crystallization, no significant variation of the diffusion coefficient with increasing concentration was observed, indicating that under such conditions up to saturation the proteins remain essentially monodisperse. Application of this technique to find crystallization conditions of other proteins is discussed.
  • References (26)
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References26
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The aggregation processes leading to crystallization and precipitation of canavalin have been investigated by dynamic light scattering (DLS) in photon correlation spectroscopy (PCS) mode. The sizes of aggregates formed under various conditions of pH, salt concentration, and protein concentrations were deduced from the correlation functions generated by the fluctuating intensity of light scattered by the solutions of the protein. Results obtained indicate that the barrier to crystallization of ca...
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Crystallization of lysozymes induced by temperature lowering has been monitored by dynamic light scattering from the onset of supersaturation to the growth of protein crystals up to a noticeable size (150–200 μm). The apparent size of the scatterers was found to increase up to a maximum value as supersaturation proceeded, then to decrease down to its initial value. Apparitions of crystals (20–30 μm) were observed during this decrease. Light scattering is thus proved to be a sensitive technique t...
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Abstract Until recently, crystallization of macromolecules was rather an empirical process. Developments in biotechnology have emphasized the need for a more rational approach. Indeed a new discipline, biological crystallogenesis, encompassing biology, physics, crystallography and related engineering aspects, is emerging. This article discusses theoretical and new methodological principles underlying biological crystallogenesis. It gives special emphasis to the concept of purity of macromolecule...
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Abstract A quick and miniature method has been devised for determining protein solubility and used to investigate the equilibrium solubility of concanavalin A from the Jack Bean with its crystals as a function of ammonium sulfate concentration, temperature and pH. The crystals were characterized by X-ray diffraction and their morphologies related to the corresponding solubilities. The protein solution concentration was estimated out of small crystallizing drops using a rapid and sensitive microa...
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