Aromatic lipophilic spin traps effectively inhibit RPE65 isomerohydrolase activity |
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Authors: | Poliakov Eugenia Parikh Toral Ayele Michael Kuo Stephanie Chander Preethi Gentleman Susan Redmond T Michael |
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Affiliation: | LRCMB, NEI, National Institutes of Health, Bethesda, Maryland 20892, United States. |
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Abstract: | We previously showed that RPE65 does not specifically produce 11-cis-retinol only but also 13-cis-retinol, supporting a carbocation or radical cation mechanism of isomerization. The intrinsic properties of conjugated polyene chains result in facile formation of radical cations in oxidative conditions. We hypothesized that such radical intermediates, if involved in the mechanism of RPE65, could be stabilized by spin traps. We tested a variety of hydrophilic and lipophilic spin traps for their ability to inhibit RPE65 isomerohydrolase activity. We found that the aromatic lipophilic spin traps such as N-tert-butyl-α-phenylnitrone (PBN), 2,2-dimethyl-4-phenyl-2H-imidazole-1-oxide (DMPIO), and nitrosobenzene (NB) strongly inhibit RPE65 isomerohydrolase activity in vitro. |
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