3-hydroxykynurenine-glucoside and 3-hydroxykynurenine

To compare the photochemical properties of UV filter molecules present in the human lens (kynurenine, KN; 3-hydroxykynurenine, 3OHKN; 3-hydroxykynurenine O-β-D-glucoside, 3OHKG; 4-(2-aminophenyl)-4-oxobutanoic acid, AHA; and glutathionyl-kynurenine, GSH-KN) with the use of the following parameters: excited singlet lifetime τ(S), fluorescence quantum yield Φ(fl), triplet quantum yield Φ(T), and photodecomposition quantum yield Φ(dec).. The excited singlet lifetimes were measured with the use of fluorescence upconversion (time resolution, 210 fs) and pump-probe transient absorption (time resolution, 200 fs) methods. The fluorescence quantum yields were determined relative to an aqueous solution of quinine bisulfate. The triplet quantum yields were measured with the use of nanosecond laser flash photolysis. The photodecomposition quantum yields were determined by steady state photolysis followed by the high-performance liquid chromatography analysis.. The secondary UV filters--AHA and GSH-KN are better photosensitizers than the primary ones--KN, 3OHKN and 3OHKG: the singlet state lifetimes of the secondary UV filters are longer, and the quantum yields of fluorescence and triplet state formation are higher.. With aging, the ratio primary/secondary UV filters in the human lens decreases from approximately 10:1 to 2:1. The obtained results demonstrate that the quality of the secondary UV filters is inferior compared to the primary ones, which may result in a higher susceptibility of old lenses to UV light. That might be an important factor for the development of the age-related cataract.

Topics: Aging; Chromatography, High Pressure Liquid; Glucosides; Glutathione; Humans; Kynurenine; Lens, Crystalline; Phenylbutyrates; Photochemistry; Photolysis; Photosensitizing Agents; Radiation-Protective Agents; Spectrometry, Fluorescence; Ultraviolet Rays

To survey the levels of protein-bound UV filters in the cortices and nuclei of normal human lenses as a function of age and to relate this to the concentration of free UV filters.. Levels of each of the three kynurenine (Kyn) UV filters, 3-hydroxykynurenine glucoside (3OHKG), Kyn, and 3-hydroxykynurenine (3OHKyn), covalently attached to proteins, were determined by using a newly developed method of reductive capture, after base treatment of the intact lens proteins.. The data show that, in the normal lens, each of the three UV filters became bound to proteins to a significant extent only after age 50 and, further, that the levels in the nucleus were much higher than in the cortex. These findings are consistent with the lens barrier that forms in middle age. 3OHKG was present at the highest levels followed by Kyn, with 3OHKyn being attached in the lowest amount. The ratio was 145:4:1 (3OHKG-Kyn-3OHKyn), with a total protein-bound UV filter concentration in the lens nucleus after age 50 of approximately 1300 picomoles/mg protein. This ratio is in agreement with 3OHKG being the most abundant free UV filter in the human lens and 3OHKyn being present in the lowest concentration with free Kyn present in intermediate amounts.. The three Kyn UV filters are bound to the nuclear proteins of all normal lenses over the age of 50. Indeed in the center of older normal lenses, the concentration of UV filters bound to proteins is approximately equal to that of the free filters. Since bound UV filters promote oxidation of proteins after exposure to wavelengths of light that penetrate the cornea, lenses in middle-aged and older individuals may be more prone to photooxidation than those of young people.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aging; Chromatography, High Pressure Liquid; Crystallins; Glucosides; Humans; Kynurenine; Lens Cortex, Crystalline; Lens Nucleus, Crystalline; Middle Aged; Protein Binding; Ultraviolet Rays

Age-dependent human lens coloration may be explained by the binding of UV filters to crystallins. It has been proposed that glutathione may compete for reaction with UV filter degradation products and therefore protect crystallins from modification. To understand this process, UV filters were quantified together with oxidized and reduced glutathione in human lenses of varying age.. Lens tissues were homogenized in ethanol to extract the UV filters. Metabolites were quantified by HPLC and correlations between them in the nuclear and cortical regions of the lens were examined.. The concentrations of the UV filters 3-hydroxykynurenine, kynurenine, and 3-hydroxykynurenine glucoside decreased linearly with age, with slightly lower levels in the nucleus than the cortex. 4-(2-Amino-3-hydroxyphenyl)-4-oxobutanoic acid glucoside was found in higher levels in the nucleus than the cortex and decreased slowly in both regions with age. Glutathionyl-3-hydroxykynurenine glucoside was present in higher concentrations in the nucleus, barely detectable in young lenses, but increased significantly after age 50. Reduced glutathione levels were lower in the nucleus and decreased in both regions with age, yet oxidized glutathione increased in the nucleus but remained constant in the cortex.. Results are consistent with a predominantly nuclear origin for both 4-(2-amino-3-hydroxyphenyl)-4-oxobutanoic acid glucoside and glutathionyl-3-hydroxykynurenine glucoside. This is in accord with their proposed mechanism of formation, which involves an initial deamination of 3-hydroxykynurenine glucoside. This process is more pronounced in older lenses, possibly because of the barrier to diffusion. The barrier may also explain the increase in nuclear oxidized glutathione that is observed with age.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aging; Chromatography, High Pressure Liquid; Glucosides; Glutathione; Humans; Kynurenine; Lens, Crystalline; Middle Aged; Phenylbutyrates; Radiation Injuries; Ultraviolet Rays