Isotachophoresis and Immunoelectrophoresis of water-soluble and -insoluble crystallins of the ageing bovine lens

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Abstract
Water-soluble (WS) and water-insoluble (WI) orystallins were prepared from calf and bovine lenses. The WS-crystallins were analyzed by capillary isotachophoresis and by Immunoelectrophoresis with polyvalent and α-, β-, γ-crystallin specific antisera. The Wl-crystallins were first solubilized in formamide and then analyzed by the same methods. The WS-crystallins, which increase in weight, but decrease in % with increasing age, show an increase of a-crystallins from 36 to 43% an increase of B-crystallins from 42 to 48% and a decrease of γ-crystallins from 22 to 9% In the WI-crystallins, which increase in weight and in % with increasing age, the α-crystallins form the main portion with an increase from 54 to 61%. The β-crystallins in WI remain fairly constant at about 37% and the γ-crystallins decrease from 8 to 2%. Quantitatively, the α-crystallin is the major constituent of the lens. The a-crystallin and the β-crystallin components are increasingly incorporated in WI with increasing age. The γ-crystallin components are selectively incorporated into WI-protein. Immunologically, α-, β- and γ-crystallins were detected by the polyvalent and specific antisera in WS- and in WI- crystallins.