Ovarian Cancer pp 645-648 | Cite as

Photoaffinity Detection of cAMP Binding Proteins in Ovarian Cancer

  • David J. Burns
  • Martin J. Hulme
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 39)

Abstract

Identification of individual cyclic AMP binding proteins in tumor cytosolic extracts is possibly owing to the ability of 32P-labeled 8-azido cyclic AMP to act as an effective analog of cAMP, to bind specifically to its protein effector sites, and on photo activation to incorporate covalently to these sites by means of a highly reactive nitrene derivative. Resolution of labeled proteins can be achieved by SDS-PAGE and visualization by autoradiography (1).

Keywords

Sucrose Magnesium Glycerol Glycine Electrophoresis 

References

  1. 1.
    Pomerantz, A. H., Rudolph, S. A., Haley, B. E., and Greengard, P. (1975) Photoaffinity labeling of a protein kinase from bovine brain with 8-azidoadenosine 3’,5’-monophosphate. Biochemistry 14, 3858–3862.CrossRefPubMedGoogle Scholar
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    Miller, W. R., Hulme, M. J., Cho-Chung, Y. S., and Elton, R. A. (1993) Types of cyclic AMP binding proteins in human breast cancers. Eur. J. Cancer 29A, 989–991.CrossRefPubMedGoogle Scholar
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    Ramage, A. D. R., Burns, D. J., and Miller, W. R. (1994) Cyclic Adenosine 3’,5’-monophosphatebinding proteins in human ovarian cancers. Br. J. Cancer 69, 186–190.CrossRefPubMedGoogle Scholar
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    Simpson, B. J. B., Ramage, A. D. R., Hulme, M. J., Burns, D. J., Katsaros, D., Langdon, S. P., et al. (1996) Cyclic adenosine 3’,5’-monophosphate-binding proteins in human ovarian cancers: correlations with clinicopathological features. Clin. Cancer Res. 2, 201–206.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2000

Authors and Affiliations

  • David J. Burns
    • 1
  • Martin J. Hulme
    • 1
  1. 1.Breast Cancer Research UnitWestern General HospitalEdinburghScotland

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