, Volume 24, Issue 1, pp 400–402 | Cite as

A new polymer supported packing material as a substitute of hydroxylapatite: Purification of Topoisomerases

  • K. Majumder
  • N. Ramesh


Hydroxylapatite (HA), a form of calcium phosphate, finds extensive usages in the fractionation and purification of proteins, enzymes and nucleic acids. However, commercial HA preparations for liquid chromatography (LC) differ from many other LC packing materials in that the former is not polymer supported. We have now developed a substitute packing material which uses Fractosil 200 as the polymer support. Other polymer supports can also be similarly used. Being Fractosil 200 supported, this Hydroxylapatite Substitute (HAS) matrix can be employed for open column LC as well as for HPLC purposes. Although the exact chemical nature of HAS is different from the conventional HA, HAS mimicks the functions of the latter support. Due to the high’ binding capacity of HA many DNA binding proteins have been partially purified on this support. In the present work HAS has been successfully used in the partial purification of Topoisomerase I and II from wheat germ.

Key Words

Liquid chromatography Hydroxylapatite substitute, HAS Protein purification Topoisomerases 

List of Common Abbreviations




Bovine serum albumin


Deoxyribonucleic acid




Ethylenediaminetetraacetic acid


40 mM Tris-acetate, 2 mM EDTA


Tris(hydroxymethyl) aminomethane


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. [1]
    A. Tiselius, S. Hjerten, O. Levin, Arch. Biochem. Biophys.,65, 132 (1956).CrossRefGoogle Scholar
  2. [2]
    G. Bernardi, Nature,206, 779 (1965).CrossRefGoogle Scholar
  3. [3]
    H. G. Martinson, Biochemistry,12, 2731 & 2737 (1973).CrossRefGoogle Scholar
  4. [4]
    G. Bernardi, in “Methods in Enzymology”,22, 325, Academic Press, New York (1971).Google Scholar
  5. [5]
    U. Schomburg, F. Grosse, Eur. J. Biochem.160, 451 (1986).CrossRefGoogle Scholar
  6. [6]
    S. Douc-Rasy, A. Kayser, J-F Riou, G. Riou, Proc. Natl. Acad. Sci. USA,83, 7152 (1986).CrossRefGoogle Scholar
  7. [7]
    K. Majumder, Ph. D. Thesis, Indian Institute of Science, Bangalore (1987).Google Scholar
  8. [8]
    For a review on phosphorylating conditions vide ref [7].Google Scholar
  9. [9]
    W. S. Dynan, J. J. Jendrisal, D. A. Hager, R. R. Burgess, J. Biol. Chem.,256, 5860 (1961).Google Scholar
  10. [10]
    D. J. H. Trafford, P. Ward, A. Ying Foo, H. L. J. Makin, J. Endocrinology,65, 44 (1975).Google Scholar
  11. [11]
    E. J. Peck, J. H. Clark, Endocrinology,101, 1034 (1977).CrossRefGoogle Scholar
  12. [12]
    R. E. Garola, W. L. McGuire, Cancer Research,37, 3329 (1977).Google Scholar
  13. [13]
    J. N. Hansen, Anal. Biochem.76, 37 (1976).CrossRefGoogle Scholar
  14. [14]
    B. Ziola, D. Scraba, Anal. Biochem.,72, 366 (1976).CrossRefGoogle Scholar
  15. [15]
    F. H. Tabak, R. A. Flavell, Necleic Acids Res.5, 2321 (1978).CrossRefGoogle Scholar

Copyright information

© Friedr. Vieweg & Sohn Verlagsgesellschaft mbH 1987

Authors and Affiliations

  • K. Majumder
    • 1
  • N. Ramesh
    • 1
  1. 1.Molecular Biophysics UnitIndian Institute of ScienceBangaloreIndia

Personalised recommendations