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Structural Studies of Human Adenine Phosphoribosyltransferase Purified by Affinity Chromatography

  • Joseph A. Holden
  • Gary S. Meredith
  • William N. Kelley
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 122B)

Abstract

Adenine phosphoribosyltransferase (APRT) catalyzes the condensation of 5-phosphoribosyl-1-pyrophosphate (PP-ribose-P) with adenine to yield adenosine 5’-monophosphate and PPi. Interest in the human enzyme has been stimulated by the findings of elevated adenine phosphoribosyltransferase activity in erythrocytes from patients with the Lesch-Nyhan syndrome1 and by the inherited deficiency of the enzyme described in a number of families.2,3,4,5 In order to better understand the nature of these alterations of adenine phosphoribosyltransferase activity at the molecular level, it is necessary to define the nature of the normal enzyme. The normal enzyme has previously been purified 33,000-fold from human erythrocytes.6 The procedure, however, is lengthy and laborious. In this report we describe a more efficient purification procedure for adenine phosphoribosyltransferase. Some of the characteristics of the highly purified enzyme, not previously reported, are described.

Keywords

Amino Acid Composition Guanidine Hydrochloride Hydrolysis Time Performic Acid Normal Enzyme 
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.

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References

  1. 1.
    Seegmiller, J.E., Rosenbloom, F.M., and Kelley, W.N. Science 155:1682–1684 (1967).PubMedCrossRefGoogle Scholar
  2. 2.
    Kelley, W.N., Levy, R.I., Rosenbloom, F.M., Henderson, J.F., and Seegmiller, J.E. J. Clin. Inves. 47:2281–2289 (1968).CrossRefGoogle Scholar
  3. 3.
    Fox, I.H., Meade, J.C., and Kelley, W.N. Amer. J. Med. 55: 614–620 (1973).PubMedCrossRefGoogle Scholar
  4. 4.
    Debray, H., Cartier, P., Temstet, A., and Cendron J. Pediat. Res. 10:762–766 (1976).PubMedGoogle Scholar
  5. 5.
    Van Acker, K.J., Simmonds, H.A., Potter, C., and Cameron, J.S. New Eng. J. Med. 297:127–132 (1977).PubMedCrossRefGoogle Scholar
  6. 6.
    Thomas, C.B., Arnold, W.J. and Kelley, W.N. J. Biol. Chem. 248:2529–2535 (1973).Google Scholar
  7. 7.
    Hughes, S.H., Wahl, G.M., and Capecchi, M.R. J. Biol. Chem. 250:120–126 (1975).Google Scholar
  8. 8.
    Holden, J.A., and Kelley, W.N. J. Biol. Chem. 253:4459–4463 (1978).Google Scholar
  9. 9.
    Simpson, R.J., Neuberger, M.R., and Liu, T.-Y. J. Biol. Chem. 251:1936–1940 (1976).Google Scholar
  10. 10.
    Hunt, L.T., and Dayhoff, M.O. Composition of Proteins in Atlas of Protein Sequence and Structure p. D-355 (1972).Google Scholar
  11. 11.
    Cohn, E.J., and Edsall, J.T. Proteins, Amino Acids and Peptides p. 370. Reinhold Publishing Corp. New York (1943).Google Scholar
  12. 12.
    Davies, G.E. and Stark, G.R. Proc. Natl. Acad. Sci. 66:651–656 (1970).PubMedCrossRefGoogle Scholar
  13. 13.
    Hirs, C.H.W. Performic Acid Oxidation in Methods in Enzymology Vol. XI, p. 197–199 (1967).Google Scholar

Copyright information

© Plenum Press, New York 1980

Authors and Affiliations

  • Joseph A. Holden
    • 1
  • Gary S. Meredith
    • 1
  • William N. Kelley
    • 1
  1. 1.Department of Internal MedicineUniversity of Michigan Medical CenterAnn ArborUSA

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