Effects of phosphorylation on binding of catecholamines to tyrosine hydroxylase: specificity and thermodynamics.
As the catalyst for the rate-limiting step in the biosynthesis of the catecholamine neurotransmitters, the activity of tyrosine hydroxylase is tightly regulated. A principle means of posttranslational regulation is reversible phosphorylation of serine residues in an N-terminal regulatory domain. Phosphorylation of serine 40 has been shown to have a large effect on the rate constant for dissociation of dopamine and a much smaller effect on that for DOPA [Ramsey, A. J., and Fitzpatrick, P. F. (1998) Biochemistry 37, 8980-8986]. To determine the structural basis for the differences in affinity and to further test the validity of the previously proposed model for regulation, the effects of phosphorylation of serine 40 on the affinities for a series of catechols have been determined. The affinities of the unphosphorylated enzyme vary by 3 orders of magnitude due to differences in the rates of dissociation. The highest affinities are found with catecholamines which lack a carboxylate. The affinities of the phosphorylated enzyme show a much smaller range. In the case of binding of dihydroxyphenylalanine, the decrease in affinity upon phosphorylation is due primarily to a decrease in the enthalpy of the interaction. Based upon these results, a structural model for the effect of phosphorylation is proposed.
Ramsey
A J
AJ
Department of Biochemistry, Department of Chemistry, Texas A&M University, College Station, Texas 77843-2128, USA.
Fitzpatrick
P F
PF
D000818:Animals; D001665:Binding Sites; D002395:Catecholamines / Q000737:chemistry* / Q000378:metabolism*; D004295:Dihydroxyphenylalanine / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D010766:Phosphorylation; D051381:Rats; D013054:Spectrophotometry, Atomic; D013056:Spectrophotometry, Ultraviolet; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D013816:Thermodynamics; D014446:Tyrosine 3-Monooxygenase / Q000737:chemistry* / Q000378:metabolism*
D002395:Catecholamines; D004295:Dihydroxyphenylalanine; D014446:Tyrosine 3-Monooxygenase