Kinetic studies on the initial crystallization process of lysozyme in the presence of D2O and H2O

In the initial stages of the crystallization of egg-white lysozyme, monomeric lysozyme aggregates rapidly and forms a nucleus in the presence of high salt concentrations. The formation process of the aggregates was examined to make clear the difference between the situations in heavy water and in water at the same sodium ion concentration. The aggregation in both cases was observed at unsaturated and/or saturated lysozyme concentrations. The turbidity at 350 nm of lysozyme increased remarkably within 60 min under each experimental condition and showed no appreciable changes over 60 min. The increase of turbidity in H2O was much slower than in D2O at the same salt concentration (3%). Lysozyme showed a critical concentration for nucleus formation whose value in H2O was lower than in D2O at 3% salt concentration. There are two different aggregation models, depending on the concentration of lysozyme. However, similar results were not obtained at 3% sodium ions in H2O. The initial aggregation rate was also dependent on the concentrations of both lysozyme and NaCl. Therefore, the effect of lysozyme concentration on the aggregation process in H2O may be smaller than in D2O.