Binding of cyclic AMP and cyclic GMP to renal cortical homogenates. Relationship with phosphorylation.
This study examined the binding of both cyclic AMP and cyclic GMP to receptor proteins in particulate and soluble subfractions of renal cortical homogenates from the golden hamster. The binding of both nucleotides was compared to subsequent effects of both nucleotides on the phosphorylation of histone from identical fractions. Cyclic AMP binding and cyclic AMP-dependent protein kinase activity predominated in the cytosol, with some binding and enzyme activity also detected in particulate fractions. Cyclic GMP and cyclic GMP-dependent protein kinase activity could only be demonstrated in cytosolic fractions and represented only 20-30% of cyclic AMP-dependent activity in this fraction. Binding of both nucleotides was highly specific, however, cyclic AMP showed some interaction with cyclic GMP binding. Evidence suggesting that each nucleotide interacts with a specific protein kinase was as follows: both the binding activity of the cyclic nucleotides and their combined protein kinase activity show additivity; cyclic AMP and cyclic GMP binding activity could be separated on sucrose gradients; cyclic AMP and cyclic GMP protein kinase activity could be separated with Sephadex G-100 chromatography, after preincubation of homogenate supernatants with either cyclic AMP or cyclic GMP. The results demonstrate the presence of both cyclic AMP- and cyclic GMP-dependent protein kinase in renal cortex.
D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism* / Q000494:pharmacology; D002373:Cyclic AMP Receptor Protein; D006152:Cyclic GMP / Q000378:metabolism* / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D007672:Kidney Cortex / Q000378:metabolism*; D008297:Male; D008647:Mesocricetus; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism
D002352:Carrier Proteins; D002373:Cyclic AMP Receptor Protein; D047908:Intracellular Signaling Peptides and Proteins; C022994:cyclic GMP-binding protein; D000242:Cyclic AMP; D011494:Protein Kinases; D006152:Cyclic GMP