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  • Title: A study of adenosine 3'-5' cyclic monophosphate binding sites of human erythrocyte membranes using 8-azidoadenosine 3'-5' cyclic monophosphate, a photoaffinity probe.
    Author: Owens JR, Haley BE.
    Journal: J Supramol Struct; 1976; 5(1):91-102. PubMed ID: 186666.
    Abstract:
    An earlier report (1a) has shown the utility of 8-N3cAMP (8-azidoadenosine-3', 5'-cyclic monophosphate) as a photoaffinity probe for cAMP binding sites in human erythrocyte membranes. The increased resolution obtained using a linear-gradient SDS polyacrylamide gel system now shows that: 1) both cAMP and 8-N3cAMP stimulate the phosphorylation by [gamma-32P]-ATP of the same red cell membrane proteins; 2) the protein of approximately 48,000 molecular weight whose phosphorylation by [gamma-32P]-ATP is stimulated by cAMP and 8-N3cAMP migrates at a slower rate than the protein in the same molecular weight range which is heavily photolabeled with [32P]-8-N3cAMP;3) other cyclic nucleotide binding sites exist besides those initially reported; 4) the variation in the ratio of incorporation of [32P]-8-N3cAMP into the two highest affinity binding sites appears to be the result of a specific proteolysis of the larger protein.
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