Cyclic AMP-dependent protein kinase is not involved in the in vivo activation of tyrosine hydroxylase in the adrenal gland after decapitation.
Tyrosine hydroxylase is activated in the adrenal gland in vivo after acute stresses, such as decapitation or electroconvulsive shock. In nonstressed animals that are anesthetized with pentobarbital prior to surgical removal of the adrenals, approximately 5-10% of the enzyme molecules are in the activated form, whereas in stressed animals, approximately 40-50% of the enzyme molecules are in the activated form. In the present study, we have tested the hypothesis that the stress-induced activation of the adrenal enzyme in vivo is due to the phosphorylation of the enzyme by cyclic AMP-dependent protein kinase. Soluble adrenal tyrosine hydroxylase prepared from either stressed or nonstressed rats is incubated in vitro with [gamma-32P]ATP and purified cyclic AMP-dependent protein kinase under optimal conditions for the phosphorylation of the enzyme. Using this assay, we have measured the number of vacant sites remaining on the enzyme, which are available for in vitro phosphorylation by cyclic AMP-dependent protein kinase. These studies suggest that the initial, in vitro rate of phosphorylation of tyrosine hydroxylase isolated from stressed rats is less than the initial rate of phosphorylation of the enzyme isolated from nonstressed rats. However, there is no significant difference in the final level of 32P phosphorylation of tyrosine hydroxylase isolated from either stressed or nonstressed rats. We conclude that, even though phosphorylation of tyrosine hydroxylase by cyclic AMP-dependent protein kinase leads to the activation of the enzyme under in vitro conditions, this mechanism cannot account for the activation of the enzyme in vivo in the adrenal gland following decapitation.
D000310:Adrenal Gland Neoplasms / Q000201:enzymology; D000311:Adrenal Glands / Q000201:enzymology*; D000818:Animals; D004789:Enzyme Activation; D007700:Kinetics; D008297:Male; D010673:Pheochromocytoma / Q000201:enzymology; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013312:Stress, Physiological / Q000503:physiopathology; D014446:Tyrosine 3-Monooxygenase / Q000302:isolation & purification / Q000378:metabolism*
D014446:Tyrosine 3-Monooxygenase; D011494:Protein Kinases