Studies on biosynthesis of soluble lens crystallin antigens in the chick by isoelectric focusing in thin-layer polyacrylamide gel.

Abstract

Abstract Biosynthesis of soluble lens crystallin antigens from 6-day embryonic to 21-day post-hatch chick was investigated by isoelectric focusing in thin-layer polyacrylamide gel. Immuno-isoelectric focusing was also performed with all the embryonic and post-hatch lens extracts against chick lens total protein antiserum in order to correlate biosynthesis and immunological detection of the antigens. From isoelectric focusing it appears that each one of the major crystallins, namely, the α-, the β-, and the delta-crystallin is heterogeneous and made up of components with different isoelectric points which are immunologically identical. Rate of synthesis of these crystallins is very high in early stages. It gradually slows down and in post-hatch lens it is continued at a very reduced rate. Radioactive profile shows that none of the crystallin components resolved by isoelectric focusing are post-synthetic derivatives of some other crystallin. We also observed that there is no synthesis of any new component in post-hatch lens. We also found that the amount of α-crystallin increased, that of delta decreases, while the β-crystallin does not show any significant difference from embryonic to post-hatch lens. Overlapping of the precipitin lines of the crystallin antigens as they appear here after immuno-isoelectric focusing is due to a prolonged period of immunodiffusion, and we have verified this with monospecific antiserum allowing less time for immunodiffusion after isoelectric focusing.