VOPROSY MEDITSINSKOI KHIMII (ISSN 0042-8809)

Method of measuring binding constants and the number of specific cyclic AMP receptor sites in tissue extracts

   
Gorbachevskaia L.V.
PubMed Id: 664475
Year: 1978  Volume: 24  Issue: 3  Pages: 423-427
A method for evaluation of binding constants and of amount of cAMP binding sites in crude tissue extracts was developed. The method is based on equilibrium binding of 3H-cAMP by proteins with subsequent ultrafiltration. Hydrolysis of cAMP and its unspecific sorption by proteins were eliminated under the conditions selected. Rat spleen cytosole contained 3.57 +/- 0.34 pmol of cAMP binding sites per mg of protein with dissociation constant of protein-cAMP complex (1.68 +/- 0.28).10(-8) M. As shown by studies on kinetics, binding constants and specificity of binding, the method permitted to evaluate quantitatively cAMP-dependent protein kinases in crude tissue extracts and to estimate their affinity to cAMP.
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Gorbachevskaia, L. V. (1978). Method of measuring binding constants and the number of specific cyclic AMP receptor sites in tissue extracts. Voprosy Meditsinskoi Khimii, 24(3), 423-427.
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