Abstract
In chaperoning dithiothreitol-denatured α-lactabumin, α-crystallin forms a chaperoning complex. In order to study the kinetics of such chaperoning it needs to be established whether the formation of the chaperoning complex is a reversible or irreversible process. The chaperoning reaction was studied by dynamic light scattering as a function of concentration and weight ratio of α-lactalbumin/α-crystallin. HPLC and subsequent SDS-PAGE gel electrophoresis experiments established that the chaperoning complex formed contains both α-crystallin and α-lactalbumin. Upon rechromatographing the chaperoning complex, the presence of monomeric α-lactalbumin has been demonstrated in addition to the chaperoning complex itself. This and equilibrium dialysis experiments demonstrated conclusively the existence of an equilibrium between monomeric partially denatured α-lactalbumin and the chaperoning complex made of α-lactalbumin and α-crystallin.
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