Tyrosine hydroxylase activity is regulated by two distinct dopamine-binding sites.
Tyrosine hydroxylase (TH), the rate-limiting enzyme in the biosynthesis of the catecholamines dopamine, noradrenaline and adrenaline, is regulated acutely by feedback inhibition by the catecholamines and relief of this inhibition by phosphorylation of serine 40 (Ser40). Phosphorylation of serine 40 abolishes the binding of dopamine to a high affinity (K(D) < 4 nM) site on TH, thereby increasing the activity of the enzyme. We have found that TH also contains a second low affinity (K(D) = 90 nM) dopamine-binding site, which is present in both the non-phosphorylated and the Ser40-phosphorylated forms of the enzyme. Binding of dopamine to the high-affinity site decreases V(max) and increases the K(m) for the cofactor tetrahydrobiopterin, while binding of dopamine to the low-affinity site regulates TH activity by increasing the K(m) for tetrahydrobiopterin. Kinetic analysis indicates that both sites are present in each of the four human TH isoforms. Dissociation of dopamine from the low-affinity site increases TH activity 12-fold for the non-phosphorylated enzyme and 9-fold for the Ser40-phosphorylated enzyme. The low-affinity dopamine-binding site has the potential to be the primary mechanism responsible for the regulation of catecholamine synthesis under most conditions.
School of Biomedical Sciences and The Hunter Medical Research Institute, Faculty of Health, The University of Newcastle, Callaghan, New South Wales, Australia.
Sarah L
SL
Gordon
Noelene S
NS
Quinsey
Peter R
PR
Dunkley
Phillip W
PW
Dickson
D000818:Animals; D001665:Binding Sites / Q000502:physiology; D002395:Catecholamines / Q000096:biosynthesis; D004298:Dopamine / Q000378:metabolism / Q000502:physiology*; D004789:Enzyme Activation / Q000502:physiology; D006801:Humans; D010766:Phosphorylation; D051381:Rats; D012694:Serine / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism*
D002395:Catecholamines; D012694:Serine; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine