Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1982 Aug 1;205(2):339–344. doi: 10.1042/bj2050339

Does superoxide anion participate in 2-oxoglutarate-dependent hydroxylation?

Elisabeth Holme 1, Göran Lindstedt 1, Sven Lindstedt 1, Ingalill Nordin 1
PMCID: PMC1158486  PMID: 6291507

Abstract

The possible role of superoxide anion in 2-oxoglutarate-coupled dioxygenase reactions has been investigated. γ-Butyrobetaine hydroxylase (EC 1.14.11.1) was inhibited by human erythrocyte superoxide dismutase (EC 1.15.1.1), probably due to release of Cu2+ or Zn2+, as the inhibition was more pronounced after heat-inactivation of the dismutase and as Cu2+ was a potent inhibitor. Bovine superoxide dismutase and the Mn2+-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. γ-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 γ-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.

Full text

PDF
339

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bankel L., Lindstedt G., Lindstedt S. Thymidine 2'-hydroxylation in Neurospora crassa. J Biol Chem. 1972 Oct 10;247(19):6128–6134. [PubMed] [Google Scholar]
  2. Bhatnagar R. S., Liu T. Z. Evidence for free radical involvement in the hydroxylation of proline: inhibition by nitro blue tetrazolium. FEBS Lett. 1972 Oct 1;26(1):32–34. doi: 10.1016/0014-5793(72)80535-0. [DOI] [PubMed] [Google Scholar]
  3. Brigelius R., Spöttl R., Bors W., Lengfelder E., Saran M., Weser U. Superoxide dismutase activity of low molecular weight Cu2 plus-chelates studied by pulse radiolysis. FEBS Lett. 1974 Oct 1;47(1):72–75. doi: 10.1016/0014-5793(74)80428-x. [DOI] [PubMed] [Google Scholar]
  4. Holme E., Lindstedt G., Lindstedt S., Tofft M. 18-O studies of the 2-ketoglutarate-dependent sequential oxygenation of thymine to 5-carboxyuracil. J Biol Chem. 1971 May 25;246(10):3314–3319. [PubMed] [Google Scholar]
  5. Holme E., Lindstedt G., Lindstedt S., Tofft M. 7-Hydroxylation of thymine in a Neurospora strain coupled to oxidative decarboxylation of 2-ketoglutarate. Biochim Biophys Acta. 1970 Jul 15;212(1):50–57. doi: 10.1016/0005-2744(70)90177-4. [DOI] [PubMed] [Google Scholar]
  6. Holme E., Lindstedt G., Lindstedt S., Tofft M. 7-Hydroxylation of thymine in a Neurospora strain coupled to oxidative decarboxylation of 2-ketoglutarate. Biochim Biophys Acta. 1970 Jul 15;212(1):50–57. doi: 10.1016/0005-2744(70)90177-4. [DOI] [PubMed] [Google Scholar]
  7. Keele B. B., Jr, McCord J. M., Fridovich I. Superoxide dismutase from escherichia coli B. A new manganese-containing enzyme. J Biol Chem. 1970 Nov 25;245(22):6176–6181. [PubMed] [Google Scholar]
  8. Kido T., Soda K. Properties of 2-nitropropane dioxygenase of Hansenula mrakii. Formation and participation of superoxide. J Biol Chem. 1978 Jan 10;253(1):226–232. [PubMed] [Google Scholar]
  9. Lindstedt G. Hydroxylation of gamma-butyrobetaine to carnitine in rat liver. Biochemistry. 1967 May;6(5):1271–1282. doi: 10.1021/bi00857a007. [DOI] [PubMed] [Google Scholar]
  10. Lindstedt G., Lindstedt S., Nordin I. Purification and properties of gamma-butyrobetaine hydroxylase from Pseudomonas sp AK 1. Biochemistry. 1977 May 17;16(10):2181–2188. doi: 10.1021/bi00629a022. [DOI] [PubMed] [Google Scholar]
  11. McCord J. M., Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem. 1969 Nov 25;244(22):6049–6055. [PubMed] [Google Scholar]
  12. Murray J. C., Cassell R. H., Pinnell S. R. Inhibition of lysyl hydroxylase by catechol analogs. Biochim Biophys Acta. 1977 Mar 15;481(1):63–70. doi: 10.1016/0005-2744(77)90137-1. [DOI] [PubMed] [Google Scholar]
  13. Myllylä R., Kuutti-Savolainen E. R., Kivirikko K. I. The role of ascorbate in the prolyl hydroxylase reaction. Biochem Biophys Res Commun. 1978 Jul 28;83(2):441–448. doi: 10.1016/0006-291x(78)91010-0. [DOI] [PubMed] [Google Scholar]
  14. Myllylä R., Schubotz L. M., Weser U., Kivirikko K. I. Involvement of superoxide in the prolyl and lysyl hydroxylase reactions. Biochem Biophys Res Commun. 1979 Jul 12;89(1):98–102. doi: 10.1016/0006-291x(79)90948-3. [DOI] [PubMed] [Google Scholar]
  15. Ohnishi T., Hirata F., Hayaish O. Indoleamine 2,3-dioxygenase. Potassium superoxide as substrate. J Biol Chem. 1977 Jul 10;252(13):4643–4647. [PubMed] [Google Scholar]
  16. Ryhänen L. Lysyl hydroxylase. Further purification and characterization of the enzyme from chick embryos and chick embryo cartilage. Biochim Biophys Acta. 1976 Jun 7;438(1):71–89. doi: 10.1016/0005-2744(76)90224-2. [DOI] [PubMed] [Google Scholar]
  17. Taniguchi T., Sono M., Hirata F., Hayaishi O., Tamura M., Hayashi K., Iizuka T., Ishimura Y. Indoleamine 2,3-dioxygenase. Kinetic studies on the binding of superoxide anion and molecular oxygen to enzyme. J Biol Chem. 1979 May 10;254(9):3288–3294. [PubMed] [Google Scholar]
  18. Tuderman L., Myllylä R., Kivirikko K. I. Mechanism of the prolyl hydroxylase reaction. 1. Role of co-substrates. Eur J Biochem. 1977 Nov 1;80(2):341–348. doi: 10.1111/j.1432-1033.1977.tb11888.x. [DOI] [PubMed] [Google Scholar]
  19. deAlvare L. R., Goda K., Kimura T. Mechanism of superoxide anion scavenging reaction by bis-(salicylato)-copper (II) complex. Biochem Biophys Res Commun. 1976 Apr 5;69(3):687–694. doi: 10.1016/0006-291x(76)90930-x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES