title: Liquid-liquid phase separation and static light scattering of concentrated ternary mixtures of bovine alpha and gammaB crystallins.
abstract: We have used light scattering, turbidimetry, and thermodynamic analysis to study the phase diagram of concentrated aqueous mixtures of the bovine lens proteins, gammaB crystallin, and alpha crystallin. We find that dilute alpha crystallin raises the phase separation temperature of concentrated gammaB crystallin, while more concentrated alpha crystallin suppresses phase separation. Very concentrated alpha/gammaB mixtures can reversibly cloud above 37 degrees C, even though gammaB alone phase separates only below temperatures near 0 degrees C, and alpha does not phase separate. At the scattering vector magnitude used, high-concentration alpha/gammaB mixtures scatter less light than the weighted average of their component alpha and gammaB solutions, while low-concentration alpha/gammaB mixtures scatter more than such a weighted average. We use a mean-field thermodynamic analysis of such ternary mixtures to show that the observed light scattering and phase boundaries of alpha and gammaB crystallin mixtures give evidence for prominent local fluctuations of relative protein composition. In the single phase, these fluctuations scatter comparatively little light, but are associated with enhanced thermodynamic instability. By applying this analysis to the experimental tie lines we estimate the magnitude of the saddlelike component of the free energy near the aqueous-gammaB critical point.
authors.affiliation: Department of Physics, Rochester Institute of Technology, Rochester, New York 14623, USA. george.thurston@rit.edu
authors.forename: George M
authors.initials: GM
authors.lastname: Thurston
mesh_terms: D000818:Animals; D002417:Cattle; D008027:Light; D044367:Phase Transition; D012542:Scattering, Radiation; D013816:Thermodynamics; D038201:alpha-Crystallins / Q000737:chemistry*; D038222:gamma-Crystallins / Q000737:chemistry*
chemical_list: D038201:alpha-Crystallins; C107738:crystallin, gammaB; D038222:gamma-Crystallins