title: On the question of translocation of heart cAMP-dependent protein kinase.
abstract: Rat hearts were perfused with epinephrine and/or 1-methyl-3-isobutylxanthine for 2 min. These agents raised the concentration of cAMP and increased the fraction of cAMP-dependent protein kinase (EC 2.7.1.70) in the active form. However, the content of cAMP-dependent protein kinase in the soluble fraction of homogenates of these hearts was reduced and the amount in the particulate fraction was increased. A similar redistribution was obtained by adding cAMP to homogenates of control hearts. The reduction in soluble protein kinase content was due to apparent binding of the free catalytic subunit of the enzyme to particulate material (12,000 times g pellet) in media of low ionic strength (smaller than 100 mM KCl). The amount bound was, therefore, proportional to the dissociation of the holoenzyme. The binding was not altered by prior boiling or trypsin treatment of the particulate material, but it was prevented or reversed by the addition of 150 mM KCl. The catalytic subunit of the protein kinase from heart also bound to particulate fractions from liver or Escherichia coli and to various denatured proteins. These findings suggest that the protein kinase activity of membranes and particulate fractions has frequently been overestimated, since isolation of particulate materials has usually been carried out at low ionic strength. The data also imply that intracellular translocation of the protein kinase catalytic subunit, at least in heart tissue, is of questionable physiological significance.
mesh_terms: D000818:Animals; D001665:Binding Sites; D000242:Cyclic AMP / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D004837:Epinephrine / Q000494:pharmacology; D004926:Escherichia coli / Q000378:metabolism; D006321:Heart / Q000187:drug effects; D008099:Liver / Q000378:metabolism; D008297:Male; D009206:Myocardium / Q000201:enzymology*; D009994:Osmolar Concentration; D010477:Perfusion; D011189:Potassium Chloride / Q000494:pharmacology; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D011955:Receptors, Drug; D013347:Subcellular Fractions / Q000201:enzymology; D014970:Xanthines / Q000494:pharmacology
chemical_list: D011955:Receptors, Drug; D014970:Xanthines; D011189:Potassium Chloride; D000242:Cyclic AMP; D011494:Protein Kinases; D004837:Epinephrine