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

Autor:	L V, Gorbachevskaia
Rok vydání:	1978
Předmět:	Cytosol Methods Animals In Vitro Techniques Receptors Cyclic AMP Spleen Rats
Zdroj:	Voprosy meditsinskoi khimii. 24(3)
ISSN:	0042-8809
Popis:	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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=pmid________::286df7e71e803548d05d20999d0f1fcd https://pubmed.ncbi.nlm.nih.gov/664475 Zobrazit plný text záznamu