Purpose: To compare the non-enzymatic glycation of different human fetal gamma-crystallins induced by glucose and fructose.
Methods: gamma 1- and gamma 2-crystallin were separated by the Sephadex gel chromatography from human fetal lenses. The non-enzymatic glycations of these gamma-crystallins were conducted by incubating with glucose or fructose at 37 degrees C for 20 days. The opacity and pellet formation as well as changes on SDS-polyacrymide gel electrophoresis (SDS-PAGE) pattern and blue fluorescence production were observed.
Results: Although the two gamma-crystallins displayed similar alterations, gamma 1-crystallin was easier to produce aggregation and insolublization in early stage (i.e. 3 days). SDS-PAGE showed that there were aggregates, formed through disulfide and non-disulfide cross-links, and degraded peptides produced. The blue fluorescence increased in glycated gamma 3-crystalline solution. Fructose had more significant effects (i.e. aggregation, degradation, producing blue fluorescence) on gamma-crystallins as compared with glucose.
Conclusion: Human fetal gamma 1- and gamma 3-crystallins are sensitive, at different extent (gamma 1 > gamma 3), to response to non-enzymatic glycation, and fructose has stronger effect than glucose on the glycation.