Advanced glycation endproducts induce photocrosslinking and oxidation of bovine lens proteins through type-I mechanism.
Advanced glycation endproducts (AGEs) have been suggested as photosensitizers that are capable of mediating eye lens photo-damage during aging. In the present work, we investigate the photo-crosslinking and oxidation of bovine lens proteins sensitized by AGEs, with special regard to low oxygen conditions. A mechanistic study was conducted using different oxygen concentrations and specific additives with the aim either to scavenge or enhance Type-I or Type-II photoprocesses. Quantum yields for Trp decomposition were determined at 5%, 20% and 100% O(2), in the presence of ferricyanide and D(2)O to elucidate the mechanism of action of AGEs. Type-I mechanism proved to be the most efficient pathway for AGE-sensitized Trp decomposition at low oxygen concentration. Photocrosslinking of lens proteins and crystallin fractions due to Type-I interaction was observed. The influence of the oxygen concentration and additives was also studied. The results show that both Type-I mechanism and oxygen-mediated reactions contribute to protein crosslinking. Carbonyl group formation due to protein photo-oxidation was detected with Oxyblot technique. The generation of high levels of hydrogen peroxide during the irradiations was detected and attributed mainly to Type-I reactions. The results support that AGEs act preferentially as Type-I sensitizers at the low oxygen concentration found in the lens and are capable of inducing protein crosslinking, oxidation and peroxide formation.
Fuentealba
Denis
D
Laboratorio de Química Biológica, Departamento de Química Física, Facultad de Química, Pontificia Universidad Católica de Chile, Santiago, Chile.
Friguet
Bertrand
B
Silva
Eduardo
E
D000818:Animals; D002417:Cattle; D003432:Cross-Linking Reagents / Q000737:chemistry* / Q000494:pharmacology; D003459:Crystallins / Q000737:chemistry / Q000378:metabolism; D017127:Glycation End Products, Advanced / Q000737:chemistry* / Q000494:pharmacology; D007908:Lens, Crystalline / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects*; D010084:Oxidation-Reduction; D010545:Peroxides / Q000737:chemistry / Q000378:metabolism; D017319:Photosensitizing Agents / Q000737:chemistry* / Q000494:pharmacology; D014364:Tryptophan / Q000737:chemistry
D003432:Cross-Linking Reagents; D003459:Crystallins; D017127:Glycation End Products, Advanced; D010545:Peroxides; D017319:Photosensitizing Agents; D014364:Tryptophan