Static light scattering from concentrated protein solutions II: experimental test of theory for protein mixtures and weakly self-associating proteins.
Using an experimental technique recently developed in this laboratory (Fernández C. and A. P. Minton. 2008. Anal. Biochem. 381:254-257), the Rayleigh light scattering of solutions of bovine serum albumin, hen egg white ovalbumin, hen egg white ovomucoid, and binary mixtures of these three proteins was measured as a function of concentration at concentrations up to 125 g/L. The measured concentration dependence of scattering of both pure proteins and binary mixtures is accounted for nearly quantitatively by an effective hard particle model (Minton A. P. 2007. Biophys. J. 93:1321-1328) in which each protein species is represented by an equivalent hard sphere, the size of which is determined by the nature of repulsive interactions between like molecules under a given set of experimental conditions. The light scattering of solutions of chymotrypsin A was measured as a function of concentration at concentrations up to 70 g/L at pH 4.1, 5.4, and 7.2. At each pH, the measured concentration dependence is accounted for quantitatively by an effective hard particle model, according to which monomeric protein may self-associate to form an equilibrium dimer and, depending upon pH, an equilibrium pentamer or hexamer.
Fernández
Cristina
C
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
Allen P
AP
D000818:Animals; D002417:Cattle; D002645:Chickens; D002918:Chymotrypsin / Q000737:chemistry; D005260:Female; D006863:Hydrogen-Ion Concentration; D008027:Light; D008956:Models, Chemical; D010047:Ovalbumin / Q000737:chemistry*; D010059:Ovomucin / Q000737:chemistry*; D012542:Scattering, Radiation; D012710:Serum Albumin, Bovine / Q000737:chemistry*
D012710:Serum Albumin, Bovine; D010059:Ovomucin; D010047:Ovalbumin; D002918:Chymotrypsin; C117608:alpha-chymotrypsin