metallothionein and dihydroethidium

Metallothionein (MT)-III is a member of a brain-specific MT family, in contrast to MT-I and MT-II that are found in most tissues and are implicated in metal ion homeostasis and as an antioxidant. To investigate the defensive role of MT-III in terms of hydroxyl radical-induced DNA damage, we used purified human MT-III. DNA damage was detected by single-strand breaks of plasmid DNA and deoxyribose degradation. In this study, we show that MT-III is able to protect against the DNA damage induced by ferric ion-nitrilotriacetic acid and H(2)O(2), and that this protective effect is inhibited by the alkylation of the sulfhydryl groups of MT-III by treatment with EDTA and N-ethylmaleimide. MT-III was also able to efficiently remove the superoxide anion, which was generated from the xanthine/xanthine oxidase system. These results strongly suggest that MT-III is involved in the protection of reactive oxygen species-induced DNA damage, probably via direct interaction with reactive oxygen species, and that MT-III acts as a neuroprotective agent.

Topics: DNA Damage; Edetic Acid; Ethidium; Ethylmaleimide; Fluorescence; Free Radical Scavengers; Humans; Hydrogen Peroxide; Hydroxyl Radical; Metallothionein; Metallothionein 3; Nerve Tissue Proteins; Neuroprotective Agents; Nitrilotriacetic Acid; Plasmids; Recombinant Proteins