title: Tryptophan cluster protects human γD-crystallin from ultraviolet radiation-induced photoaggregation in vitro.
abstract: Exposure to ultraviolet radiation (UVR) is a significant risk factor for age-related cataract, a disease of the human lens and the most prevalent cause of blindness in the world. Cataract pathology involves protein misfolding and aggregation of the primary proteins of the lens, the crystallins. Human γD-crystallin (HγD-Crys) is a major γ-crystallin in the nucleus of the human lens. We report here analysis of UVR-induced damage to HγD-Crys in vitro. Irradiation of solutions of recombinant HγD-Crys with UVA/UVB light produced a rise in solution turbidity due to polymerization of the monomeric crystallins into higher molecular weight aggregates. A significant fraction of this polymerized protein was covalently linked. Photoaggregation of HγD-Crys required oxygen and its rate was protein concentration and UVR dose dependent. To investigate the potential roles of individual tryptophan residues in photoaggregation, triple W:F mutants of HγD-Crys were irradiated. Surprisingly, despite reducing UVR absorbing capacity, multiple W:F HγD-Crys mutant proteins photoaggregated more quickly and extensively than wild type. The results reported here are consistent with previous studies that postulated that an energy transfer mechanism between the highly conserved pairs of tryptophan residues in HγD-Crys could be protective against UVR-induced photodamage.
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge, MA.
authors.forename: Nathaniel
authors.initials: N
authors.lastname: Schafheimer
authors.forename: Jonathan
authors.initials: J
authors.lastname: King
mesh_terms: D018360:Crystallography, X-Ray; D006801:Humans; D046529:Microscopy, Electron, Transmission; D008958:Models, Molecular; D011108:Polymers / Q000737:chemistry; D014364:Tryptophan / Q000737:chemistry*; D014466:Ultraviolet Rays; D038222:gamma-Crystallins / Q000737:chemistry* / Q000528:radiation effects
chemical_list: D011108:Polymers; D038222:gamma-Crystallins; D014364:Tryptophan