Ontogeny of human lens crystallins.

The soluble proteins from prenatal and neonatal human lenses were fractionated by gel filtration into four distinct size classes viz. high molecular weight alpha-crystallin (HM-alpha), alpha-crystallin, intermediate molecular weight (IMW) proteins and low molecular weight (LMW) proteins. Extinction coefficients of the isolated proteins were determined and used to calculate the proportions of each fraction on a weight basis. The distributions of polypeptides within each of these fractions were analyzed by SDS gel electrophoresis and isoelectric focussing, followed by densitometric scanning of the gels. HM-alpha is detectable as early as the 14th week of gestation and its proportions increase rapidly, to about 9% of the total protein in the 1 year postnatal lens. The alpha-crystallin, IMW and LMW fractions show concomitant decreases and by 1 year they represent about 34, 35 and 18%, respectively. However, the proportions of IMW and LMW proteins do not accurately reflect those of the beta- and gamma-crystallins, as is often assumed. Substantial levels of non-crystallin polypeptides were found in the IMW protein fractions, including a group of very basic polypeptides (VBP) which comprised up to one-third of this material in the youngest lenses. Moreover, in postnatal lenses beta s-crystallin accounted for almost half of the LMW proteins. These points considered, alpha-crystallin is the major protein in the neonatal lens (approximately 42%, including HM-alpha), followed by the beta-crystallin (approximately 36% at most and probably less), the gamma-crystallins (approximately 11%) and beta s-crystallin (approximately 9%). Substantial changes in the proportions of specific polypeptides were observed throughout early development. These appear to result from changes at the level of protein synthesis and from postsynthetic modification. The A:B subunit ratio of alpha-crystallin drops from about 12 to below 3 during early development. This coincides with increasing levels of various deamidated and degraded subunits. The major beta-crystallin polypeptide also undergoes rapid deamidation and evidence is presented suggesting that the gamma-crystallins are subject to similar modification. The most dramatic changes were observed in the constituents of the LMW proteins. The synthesis of gamma-crystallins virtually ceases at some time around birth. At the same time, the levels of beta s-crystallin undergo an explosive increase. These and other changes are discussed in terms of their possible functional significance. They are also related to the complex protein status found in old lenses.