MONITORING LIGHT-INDUCED CHANGES IN ISOLATED, INTACT EYE LENSES

Fluorescence spectral changes occurring upon irradiation with 300 nm light have been monitored in situ in isolated, intact, whole lenses from the eyes of several species. The findings corroborate observations on other individual constituent protein molecules in the solution state, and also reveal features attributable to the supramolecular protein assembly that exists in the whole lens. Irradiation of the lens with 300 nm light causes red shifts in the tryptophan emission spectrum, suggesting alterations in the protein packing in the lens. Intermolecular energy transfer from tryptophan to one of the photoproducts, presumably N-formylkynurenine (N-FK), occurs in the condensed-phase sample. The N-FK formed is photodegraded efficiently in the lens, indicating that the photodynamic effects of endogenous N-FK might not be as severe as has been thought. Species variation in the photoevents are evident, particularly in avian lenses that contain the variant δ-crystallin as the core protein. The photoinduced changes in the near-UV circular dichroism of δ-crystallin (which is α-helical, as opposed to the β-sheet structure of α-, β-, and -γ-crystallins), isolated from chicken lenses, are remarkably different from other crystallins. Irradiation of δ-crystallin leads to a drastic reduction of circular dichroism intensity in the 250–300 nm region, whereas no changes are seen in the peptide absorption band.