The low affinity dopamine binding site on tyrosine hydroxylase: the role of the N-terminus and in situ regulation of enzyme activity.
Sarah L Gordon, Julianne K Webb, Jacqueline Shehadeh, Peter R Dunkley, Phillip W Dickson
Neurochem Res | Published : 2009
Tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis, is inhibited in vitro by catecholamines binding to two distinct sites on the enzyme. The N-terminal regulatory domain of TH contributes to dopamine binding to the high affinity site of the enzyme. We prepared an N-terminal deletion mutant of TH to examine the role of the N-terminal domain in dopamine binding to the low affinity site. Deletion of the N-terminus of TH removes the high affinity dopamine binding site, but does not affect dopamine binding to the low affinity site. The role of the low affinity site in situ was examined by incubating PC12 cells with L-DOPA to increase the cytosolic catecholamine conc..View full abstract