THE REACTION OF METHYLGLYOXAL WITH HUMAN AND BOVINE LENS PROTEINS

Methylglyoxal is an endogenous metabolite that increases in diabetes and has been implicated in some of its long-term complications such as retinopathy, neuropathy and cataract. We investigated the reaction of methylglyoxal with isolated human and bovine lens crystallins (alpha, beta(H), beta(L) and gamma). After 7 days incubation at 37 degrees C and pH 6.9, the reaction of methylglyoxal with lens proteins yielded stable adducts that exhibited fluorescent properties. SDS-polyacrylamide gel electrophoresis was used to monitor aggregation and crosslinking of the modified protein and autoradiography showed that [C-14]methylglyoxal was incorporated into all the protein bands. Bovine gamma-crystallin was the most reactive towards methylglyoxal. Reaction of methylglyoxal with bovine gamma II-crystallin, which is found mainly in the lens nucleus, could alter the charge surface network of the molecule, resulting in aggregation, increased light scattering and hence cataract. Modification of gamma II-crystallin by methylglyoxal produced an overall loss of positive charge and an increase in molecular weight and non-disulfide covalent crosslinking. Amino acid analysis of the modified gamma II-crystallin showed a loss of 47% of arginine residues.