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High oxygen tension, exposure to light, and the biochemical events of vision generate significant oxidative stress in the retina and the retinal pigment epithelium (RPE). Understanding the mechanisms and basis of susceptibility to progressive retinal diseases involving oxidative damage such as age-related macular degeneration (AMD) remains a major challenge. Here microsomal glutathione S-transferase (MGST1) is shown to be a dominant, highly expressed enzyme in bovine and mouse RPE microsomes that displays significant reduction activity toward synthetic peroxides, oxidized RPE lipids, and oxidized retinoids. This enzymatic reduction activity (GPx) can be partially neutralized with a monoclonal anti-MGST1 antibody developed in this study. MGST1-transfected HEK293 cells exhibited greater viability (70 +/- 4% survival) compared with untransfected control cells (46 +/- 4% survival) when challenged with 20 microM H(2)O(2), and greater viability of MGST1-transfected cells following challenge with oxidized docosahexaenoic acid was also observed. Cultured ARPE19 cells transfected with silencing MGST1 siRNAs exhibited lower expression of MGST1 (12% and 26% of the controls) and significantly lower GPx activity (44 +/- 13%) and, thus, were more susceptible to oxidative damage. Immunoblotting revealed that the in vivo expression of MGST1 in mouse RPE decreases 3-4-fold with age, to trace levels in 18-month-old mice. GPx activity in the RPE was also found to be reduced in 12-month-old mice to approximately 67%. These results support an important protective function for MGST1 against oxidative insult in the RPE that decreases with age and suggest that this enzyme may play a role in the development of age-related diseases such as AMD.

biochemistry

Microsomal Glutathione <i>S</i>-Transferase 1 in the Retinal Pigment Epithelium: Protection against Oxidative Stress and a Potential Role in Aging<sup>,</sup>

Microsomal Glutathione S-Transferase 1 in the Retinal Pigment Epithelium: Protection against Oxidative Stress and a Potential Role in Aging,

Microsomal Glutathione S-Transferase 1 in the Retinal Pigment Epithelium: Protection against Oxidative Stress and a Potential Role in Aging^,