The formation of oxidatively induced high-molecular-weight aggregate of alpha-/gamma-crystallins.

ABSTRACT

alpha-/gamma-Crystallin interactions under oxidation with ascorbate-FeCl3-EDTA-H2O2 followed by dialysis have been studied. A high-molecular-weight aggregate (HMWA) composed of alpha- and gamma-crystallin was observed for the mixture of the dialyzed alpha-crystallin and the oxidized gamma-crystallin through gel-filtration chromatography. This illustrates an interaction between alpha-crystallin and partially denatured gamma-crystallin induced by oxidation. No HMWA formation was observed under the condition without dialysis and/or with the addition of catalase to the oxidized gamma-crystallin prior to the addition of alpha-crystallin. More HMWA was formed by oxidized gamma-crystallin followed by the addition of alpha-crystallin than by simultaneous oxidation of both alpha- and gamma-crystallins. Conformational changes of alpha-crystallin during oxidation analyzed by circular dichroism spectra showed that oxidized alpha-crystallin can gradually be restored to an ordered structure through dialysis. The overall results imply that structural changes of both alpha- and gamma-crystallins and dialysis are required to form HMWA. The observation of this oxidatively induced chaperone/substrate complex suggests that an efficient chaperone-like protective action against oxidative insults may exist in vivo.