Affinity Labeling Steroids for Characterization of Steroid Binding Sites

  • James C. Warren
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 36)


Because enzyme active sites have been identified by affinity labeling with substrate derivatives, we presumed that a similar approach might be used to characterize macromolecular steroid-binding sites. Affinity labeling (site directed irreversible binding) depends upon attaching to the steroid a reagent group capable of reaction with amino acid residues present at a macromolecular steroid binding site. Concentration of an appropriate reagent group at the steroid binding site by the steroid moiety favors covalent bond formation at the binding site as compared to the protein molecule in general. This basic scheme is illustrated in Fig. 1, which contrasts the reversible finding of receptor and steroid with the case where the same steroid bears a reagent group (X) capable of reacting with a residue at the binding site (Y). Note that the covalent bonding step (if a true covalent bond is formed) is irreversible and the steroid is not capable of leaving the site. While some affinity labeling compounds form only pseudo-covalent bonds and will dissociate from the site, it is possible to make derivatives that will form covalent bonds which endure even acid hydrolysis of the macromolecule.


Pyruvate Kinase Amino Acid Analysis Estrogenic Activity Histidine Residue Enzyme Active Site 
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.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Chin, C.C., and Warren, J.C., J. Biol. Chem., 243; 5056, 1968.PubMedGoogle Scholar
  2. 2.
    Muldoon, T.G., and Warren, J.C., J. Biol.Chem.,2445430, 1969.PubMedGoogle Scholar
  3. 3.
    Barker, K.L., and Warren, J.C., Endocrinology, 80: 536, 1967.PubMedCrossRefGoogle Scholar
  4. 4.
    Martin, L., Steroids, 13: 1, 1969.PubMedCrossRefGoogle Scholar
  5. 5.
    Korenman, S.G., Steroids, 13: 163, 1969PubMedCrossRefGoogle Scholar
  6. 6.
    Ellis, R.W., and Warren, J.C., Steroids, 17: 331, 1971.PubMedCrossRefGoogle Scholar
  7. 7.
    Muldoon, T.C., Biochemistry10: 3780,1971.PubMedCrossRefGoogle Scholar
  8. 8.
    Ganguly, M., and Warren, J.C., J. Biol. Chem., 246: 3646, 1971.PubMedGoogle Scholar
  9. 9.
    Sweet, F., and Warren, J.C., Biochim. Biophys. Acta,260: 759, 1972.PubMedGoogle Scholar
  10. 10.
    Sweet, F., Arias, F., and Warren, J.C., J. Biol. Chem.,247: 3424, 1972.PubMedGoogle Scholar
  11. 11.
    Chin, C.C., and Warren, J.C., Biochemistry, 11: 2720, 1972.PubMedCrossRefGoogle Scholar
  12. 12.
    Betz, G., and Warren, J.C., Arch. Biochem. Biophys.,128: 745, 1968.PubMedCrossRefGoogle Scholar
  13. 13.
    Betz, G., Thesis, University of Kansas School of Medicine, Kansas City, Kansas, 1968.Google Scholar
  14. 14.
    Ellis, R.W., and Warren, J.C., Endocrinology, 88: 1136, 1971.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1973

Authors and Affiliations

  • James C. Warren
    • 1
  1. 1.Departments of Obstetrics-Gynecology and Biological ChemistryWashington University School of MedicineSt. LouisUSA

Personalised recommendations