A systematic approach to evaluate the modification of lens proteins by glycation-induced crosslinking.
To systematically evaluate the modification of lens proteins by aldose and dicarbonyl sugars during the glycation process, the sugar-dependent incorporation of Lys and Arg, SDS-PAGE profile, amino acid analysis, and fluorophore formation (excitation 370 nm/emission 440 nm) were determined. Reaction mixtures with glycolaldehyde, glyceraldehyde, threose and 3-deoxythreosone showed the greatest extent of Lys crosslinking and fluorescence formation. An increase in fluorescence intensity, but a decrease in Lys and Arg crosslinking, was found with glyoxal, methylglyoxal, hydroxypyruvaldehyde and threosone. In addition glyoxal, methylglyoxal and hydroxypyruvaldehyde caused the specific loss of Arg residues in lens proteins. Reaction mixtures with xylose, xylosone, glucose, glucosone and 3-deoxyglucosone exhibited the least protein modifications; however, incubation with 3-deoxyxylosone resulted in extensive loss of Lys and Arg residues, a higher extent of Lys or Arg crosslinking and significant fluorophore formation. Each sugar exhibited unique characteristics in the modification of lens proteins by glycation. To validly compare the protein modifications occurring during glycation reactions, a systematic approach was employed to evaluate the potential role of aldose and dicarbonyl sugars in protein modification.
Lee
K W
KW
Mason Eye Institute, University of Missouri, School of Medicine, Columbia 65212, USA.
Simpson
G
G
Ortwerth
B
B
D001120:Arginine / Q000737:chemistry; D002241:Carbohydrates / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry; D003459:Crystallins / Q000737:chemistry*; D005453:Fluorescence; D006023:Glycoproteins / Q000737:chemistry*; D008239:Lysine / Q000737:chemistry