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Y-C Chai, G Hoppe, JJ Mieyal, J Sears; Reduction of Dehydroascorbate May Depend on Glutaredoxin in Human Retinal Pigment Epithelium . Invest. Ophthalmol. Vis. Sci. 2002;43(13):711.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To determine whether catalytic reduction of dehydroascorbate in human retinal pigment epithelium (RPE) is mediated by the glutaredoxin protein. Methods: Western blot analysis of glutaredoxin expression in retinal pigment epithelium from donor eyes was performed. Bovine RPE extract and recombinant glutaredoxin was tested for dehydroascorbate reductase activity using spectrophotometry. Glutaredoxin and glutaredoxin mutant (Cys23 to Ser) cDNA were cloned into pcDNA 3.1 and transfected into RPE-J cells to determine whether glutaredoxin transfectants demonstrate an increased rate of dehydroascorbate reduction in RPE-J cells. Results: Native human RPE and bovine RPE express glutaredoxin protein. Recombinant glutaredoxin and bovine RPE extracts can reduce dehydroascorbate to ascorbate with a rate of 1.2 nmol/min/mg of protein and 1.0 pmol/min/mg of protein, respectively. Transfected RPE-J cells overexpress glutaredoxin and are being tested for the dehydroascorbate reducing activity. Conclusion: Glutaredoxin may be involved with ascorbate recycling in retinal pigment epithelium. Ascorbate regeneration may be increased by catalytic reduction of dehydroascorbate.
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