title: Does superoxide anion participate in 2-oxoglutarate-dependent hydroxylation?
abstract: The possible role of superoxide anion in 2-oxoglutarate-coupled dioxygenase reactions has been investigated. gamma-Butyrobetaine hydroxylase (EC 1.14.11.1) was inhibited by human erythrocyte superoxide dismutase (EC 1.15.1.1), probably due to release of Cu(2+) or Zn(2+), as the inhibition was more pronounced after heat-inactivation of the dismutase and as Cu(2+) was a potent inhibitor. Bovine superoxide dismutase and the Mn(2+)-containing superoxide dismutase from Escherichia coli were not inhibitory. Superoxide anion generated from xanthine/xanthine oxidase was not stimulatory and could not replace ascorbate. Thymine 7-hydroxylase (EC 1.14.11.6) and thymidine 2'-hydroxylase (EC 1.14.11.3) were not inhibited by erythrocyte superoxide dismutase or stimulated by superoxide anion. gamma-Butyrobetaine hydroxylase was inhibited by a number of low-molecular-weight compounds, such as tetranitromethane, Nitro Blue Tetrazolium, adrenaline and Tiron, which may act as scavengers of superoxide anion. Involvement of this radical in other oxygenase reactions has been inferred from the findings that they were inhibitory for the respective enzymes. Several of these compounds also inhibited gamma-butyrobetaine hydroxylase. It could be concluded from these experiments, however, that mechanisms other than disposal of superoxide anion might equally well be operative, such as hydrophobic interaction with the enzyme protein and interaction with compounds required for full enzymic activity, e.g. iron and ascorbate. The results appear to rule out a requirement for superoxide anion generated in free solution, and have not yielded evidence for participation of enzyme-bound superoxide anion in 2-oxoglutarate-dependent hydroxylations.
mesh_terms: D003300:Copper / Q000378:metabolism; D006873:Hydroquinones / Q000378:metabolism; D006900:Hydroxylation; D007656:Ketoglutaric Acids / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D009580:Nitroblue Tetrazolium / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D011809:Quinones / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism*; D013774:Tetranitromethane / Q000378:metabolism; D050563:gamma-Butyrobetaine Dioxygenase
chemical_list: D006873:Hydroquinones; D007656:Ketoglutaric Acids; D011809:Quinones; D013481:Superoxides; D009580:Nitroblue Tetrazolium; D003300:Copper; D006899:Mixed Function Oxygenases; D050563:gamma-Butyrobetaine Dioxygenase; D013482:Superoxide Dismutase; D013774:Tetranitromethane; D010100:Oxygen