Ascorbate free radical reductase and ascorbate redox cycle in the human lens.
The presence of ascorbate free radical (AFR) reductase (NADH:AFR oxidoreductase, EC 1.6.5.4) in senile cataractous human lenses was demonstrated by measuring spectrophotometrically NADH oxidation in the presence of ascorbate plus ascorbate oxidase. About 80-85% of the lens AFR reductase was probably recovered in the supernatant of the lens homogenate. Michaelis constants of the reductase were about 10 microM and less than 1 microM for AFR and NADH, respectively. We also showed that AFR reductase activities in the cataractous lenses tended to decrease with increase of insoluble lens protein contents, or showed rather the possibility that the reductase activity may have decreased before the lens protein aggregation. In the highest activity group (about 120-160 nmol NADH oxidized/min/lens), it was roughly calculated that the reductase in the lens could re-reduce immediately the total (or almost total) amount of AFR produced there by ascorbate oxidation even at a high rate of 600-800 microM/min, if NADH concentration in the lens were sufficiently maintained. The above results suggested that AFR reductase in the human lens plays important roles in ascorbate regeneration of its redox cycle, and that activity loss of AFR reductase, as well as of superoxide dismutase, glutathione peroxidase and glutathione reductase, may be responsible for the oxidative changes in lens proteins with the development of senile cataracts.
Bando
M
M
Department of Ophthalmology, Tokai University School of Medicine, Isehara, Japan.
Obazawa
H
H
D000368:Aged; D000369:Aged, 80 and over; D001205:Ascorbic Acid / Q000378:metabolism*; D002386:Cataract / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D007908:Lens, Crystalline / Q000032:analysis / Q000201:enzymology* / Q000378:metabolism; D009243:NAD / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D010084:Oxidation-Reduction; D014020:Tissue Extracts / Q000494:pharmacology
D014020:Tissue Extracts; D009243:NAD; D009247:NADH, NADPH Oxidoreductases; C023927:monodehydroascorbate reductase (NADH); D001205:Ascorbic Acid