Miniaturized scintillation technique for protein solubility determinations

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Abstract
We have developed a miniaturized (volume of crystallizing solution ∼100 μl) technique for the determination of proteinsolubility as a function of temperature. After nucleation, crystals are detected by the light they scatter. Then the temperature at which a solution with the initial concentration is in equilibrium with the crystals is sought by stepwise, equilibrium dissolution of the crystals. The approach to solubility from the side of dissolution provides for higher accuracy of the determinations. The method was used to determine the temperature dependence of the solubility of human hemoglobin (Hb) C, for which high-resolution x-ray crystallography data are needed to understand the structural basis for the drastically different in vivo aggregation/crystallization behavior of β 6 Hb mutants.