Static light scattering from concentrated protein solutions, I: General theory for protein mixtures and application to self-associating proteins.
Exact expressions for the static light scattering of a solution containing up to three species of point-scattering solutes in highly nonideal solutions at arbitrary concentration are obtained from multicomponent scattering theory. Explicit expressions for thermodynamic interaction between solute molecules, required to evaluate the scattering relations, are obtained using an equivalent hard particle approximation similar to that employed earlier to interpret scattering of a single protein species at high concentration. The dependence of scattering intensity upon total protein concentration is calculated for mixtures of nonassociating proteins and for a single self-associating protein over a range of concentrations up to 200 g/l. An approximate semiempirical analysis of the concentration dependence of scattering intensity is proposed, according to which the contribution of thermodynamic interaction to scattering intensity is modeled as that of a single average hard spherical species. Simulated data containing pseudo-noise comparable in magnitude to actual experimental uncertainty are modeled using relations obtained from the proposed semiempirical analysis. It is shown that by using these relations one can extract from the data reasonably reliable information about underlying weak associations that are manifested only at very high total protein concentration.
Minton
Allen P
AP
Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, U.S. Department of Health and Human Services, Bethesda, Maryland, USA. minton@helix.nih.gov
D008027:Light; D008962:Models, Theoretical; D011506:Proteins / Q000737:chemistry*; D012542:Scattering, Radiation; D012996:Solutions
D011506:Proteins; D012996:Solutions