Deamidation and disulfide bonding in human lens γ-crystallins

Detailed analysis of the three gamma-crystallins present in the water-soluble portion of human lenses, gamma S, gamma D and gamma C, has identified disulfide bonding and deamidation as the major post-translational modifications of these crystallins. Chromatographic and mass spectrometric techniques were used to isolate and identify water-soluble gamma-crystallins from normal lenses, ages 32 week gestation, 0 day old, 4 day old, 19, 31, 45 and 55 year old. The amino acid sequences of the gamma-crystallins were confirmed and/or corrected by mass spectrometric analysis of peptides produced by enzymatic digestion or chemical fragmentation of the isolated crystallins. The molecular weight of peptides were also used to identify, locate and quantify modifications. Each of the gamma-crystallins had two disulfide bonds as well as several deamidated glutamine and asparagine residues. The extent of disulfide bond formation and deamidation appeared to increase with the age of the lens. This examination of normal human lens gamma-crystallins, the first detailed characterization of the gamma-crystallins, will provide a basis fbr comparison with modifications found in the water-insoluble portion and in cataractous lenses. (C) 1998 Academic Press.