Calcium, ATP, and magnesium activate soluble tyrosine hydroxylase from rat striatum.
Soluble tyrosine hydroxylase (TH) prepared from rat striatum by sonication, centrifugation, and gel filtration on Sephadex G-25 was activated by preincubation with Ca2+, ATP, and Mg2+. Activation occurred with micromolar concentrations of Ca2+ and required the presence of both ATP and Mg2+. The activation was reversible and was characterized by a large decrease of apparent Km for the pteridine cofactor and a small increase of Vmax. Ca2+-induced activation was small when TH activity was assayed at pH values near the optimum, but the magnitude of the activation increased with increasing assay pH. The activation apparently did not involve Ca2+-activated protease because it was not affected by the protease inhibitor leupeptin. Nor did it involve cyclic AMP-dependent protein kinase, as evidenced by the failure of the heat-stable inhibitor of this kinase to decrease Ca2+-induced TH activation. Furthermore, the activation of TH evoked by Ca2+ and that produced by cyclic AMP was additive. These experiments indicate that striatal TH can be activated in vitro by an endogenous Ca2+-dependent mechanism. The similarity of the Ca2+-induced activation of TH to that elicited by increased neuronal activity and terminal depolarization is discussed.
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002154:Calpain; D003342:Corpus Striatum / Q000201:enzymology*; D000242:Cyclic AMP / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology*; D008297:Male; D011480:Protease Inhibitors; D047428:Protein Kinase Inhibitors; D011622:Pterins / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism*
D011480:Protease Inhibitors; D047428:Protein Kinase Inhibitors; D011622:Pterins; D000255:Adenosine Triphosphate; C017523:6-methyltetrahydropterin; D000242:Cyclic AMP; D014446:Tyrosine 3-Monooxygenase; D002154:Calpain; D008274:Magnesium; D002118:Calcium