Negative second virial coefficients as predictors of protein crystal growth: evidence from sedimentation equilibrium studies that refutes the designation of those light scattering parameters as osmotic virial coefficients.
The effects of ammonium sulphate concentration on the osmotic second virial coefficient (BAA/MA) for equine serum albumin (pH 5.6, 20 degrees C) have been examined by sedimentation equilibrium. After an initial steep decrease with increasing ammonium sulphate concentration, BAA/MA assumes an essentially concentration-independent magnitude of 8-9 ml/g. Such behaviour conforms with the statistical-mechanical prediction that a sufficient increase in ionic strength should effectively eliminate the contributions of charge interactions to BAA/MA but have no effect on the covolume contribution (8.4 ml/g for serum albumin). A similar situation is shown to apply to published sedimentation equilibrium data for lysozyme (pH 4.5). Although termed osmotic second virial coefficients and designated as such (B22), the negative values obtained in published light scattering studies of both systems have been described incorrectly because of the concomitant inclusion of the protein-salt contribution to thermodynamic nonideality of the protein. Those negative values are still valid predictors of conditions conducive to crystal growth inasmuch as they do reflect situations in which there is net attraction between protein molecules. However, the source of attraction responsible for the negative virial coefficient stems from the protein-salt rather than the protein-protein contribution, which is necessarily positive.
Deszczynski
Marcin
M
National Centre for Macromolecular Hydrodynamics, University of Nottingham, Sutton Bonington, LE12 5RD, UK.
Harding
Stephen E
SE
Winzor
Donald J
DJ
D000645:Ammonium Sulfate / Q000378:metabolism; D000818:Animals; D045424:Complex Mixtures / Q000737:chemistry / Q000302:isolation & purification; D003460:Crystallization; D006736:Horses; D008027:Light; D009997:Osmotic Pressure; D012542:Scattering, Radiation; D012709:Serum Albumin / Q000737:chemistry / Q000302:isolation & purification*; D013816:Thermodynamics; D014461:Ultracentrifugation
D045424:Complex Mixtures; D012709:Serum Albumin; D000645:Ammonium Sulfate