Advertisement

Combined Use of Radioenzymatic Assay and High Pressure Liquid Chromatography for the Detection of Myocardial Xanthine Oxidase/Dehydrogenase

  • Bob Schoutsen
  • Peter de Tombe
  • Eef Harmsen
  • Elisabeth Keijzer
  • Jan Willem de Jong
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 165)

Abstract

One of the current interests in xanthine oxidase (XO; EC 1.2.3.2; electron acceptor is O2, ref. 1) is its possible role in the initiation of atherosclerosis (refs. 2,3). To study the effects of bovine milk XO in rat heart, more knowledge is needed of the XO activity in this tissue, since data with respect to specific activity vary (refs.4–6). Several methods are available to measure XO activity (refs. 7,8). At the moment XO is thought to be present in mammalian tissues mainly as xanthine dehydrogenase (XD; EC 1.2.1.37; physiologic electron acceptor NAD+, ref. 1). The present paper describes the detection of XO and XD in rat heart by radioenzymatic assay in which the oxypurines are separated by high pressure liquid chromatography (HPLC).

Keywords

Uric Acid Xanthine Oxidase Xanthine Dehydrogenase Uric Acid Production High Pressure Liquid Chroma System 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    W.R. Waud, and K.V. Rajagopalan, The mechanism of conversion of rat liver xanthine dehydrogenase from an NAD+-dependent form (type D) to an O2-dependent form (type O), Arch. Biochem. Biophys. 172: 365–379 (1976).PubMedCrossRefGoogle Scholar
  2. 2.
    K.A. Oster, Bovine milk xanthine oxidase as one of the dietary causes of early atherosclerosis, Med. Counterpoint 6: 39–42 (1974).Google Scholar
  3. 3.
    C.J. Carr, J.M. Talbot, and K.D. Fisher, A review of the significance of bovine milk xanthine oxidase in the etiology of atherosclerosis, review Food and Drug Adm., F.A.S.E.B. Life Sci. Res. Office, Bethesda, pp 1–65 (1975).Google Scholar
  4. 4.
    M.H. Maguire, M.C. Lukas, and J.F. Rettie, Adenine nucleotide salvage synthesis in the rat heart; pathways of adenosine salvage, Biochim. Biophys. Acta 262: 108–115 (1972).PubMedCrossRefGoogle Scholar
  5. 5.
    C.R.H. Ramboer, A sensitive and nonradioactive assay for serum and tissue xanthine oxidase, J. Lab. Clin. Med. 74: 828–835 (1969).PubMedGoogle Scholar
  6. 6.
    M.P.S. Gandhi, and S.P. Ahuja, Absorption of xanthine oxidase from the intestines of rats and rabbits and its role in initiation of atherosclerosis, Z. Vet. Med. A. 26: 635–642 (1979).Google Scholar
  7. 7.
    T.M. Dougherty, A sensitive assay for xanthine oxidase using commercially available (14C)xanthine, Anal. Biochem. 74: 604–608 (1976).PubMedCrossRefGoogle Scholar
  8. 8.
    G.J. Putterman, B. Shaikh, M.R. Hallmark, C.G. Sawyer, C.V. Hixson, and F. Perini, Simultaneous analysis of substrates, products, and inhibitors of xanthine oxidase by high-pressure liquid chromatography and gas chromatography, Anal. Biochem. 98: 18–26 (1979).PubMedCrossRefGoogle Scholar
  9. 9.
    H. Stam, and J.W. de Jong, Sephadex-induced reduction of coronary flow in the isolated rat heart: a model for ischemic heart disease, J. Mol. Cell. Cardiol. 9: 633–650 (1977).PubMedCrossRefGoogle Scholar
  10. 10.
    M.M. Jezewska, and Z.W. Kaminski, Xanthine oxidoreductase inhibition by NADH as a regulatory factor of purine metabolism, Adv. Exp. Med. Biol. 122B: 35–40 (1980).CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1984

Authors and Affiliations

  • Bob Schoutsen
    • 1
  • Peter de Tombe
    • 1
  • Eef Harmsen
    • 1
  • Elisabeth Keijzer
    • 1
  • Jan Willem de Jong
    • 1
  1. 1.Cardiochemical Laboratory, Thoraxcenter, ErasmusUniversity RotterdamRotterdamThe Netherlands

Personalised recommendations