Diagnostic of precipitant for biomacromolecule crystallization by quasi-elastic light-scattering.
The translational diffusion coefficient D25,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 diffusion coefficient of the scatters 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 D25,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.
Mikol
V
V
Laboratoire de Biochimie, Institut de Biologie Moléculaire et Cellulaire, Strasbourg, France.
Hirsch
E
E
Giegé
R
R
D000645:Ammonium Sulfate; D011232:Chemical Precipitation; D003208:Concanavalin A; D003460:Crystallization; D006863:Hydrogen-Ion Concentration; D008027:Light; D046911:Macromolecular Substances; D009113:Muramidase; D009994:Osmolar Concentration; D012542:Scattering, Radiation; D012965:Sodium Chloride
D046911:Macromolecular Substances; D003208:Concanavalin A; D012965:Sodium Chloride; D009113:Muramidase; D000645:Ammonium Sulfate