The Dopamine/D1 receptor mediates the phosphorylation and inactivation of the protein tyrosine phosphatase STEP via a PKA-dependent pathway.
The striatal-enriched protein tyrosine phosphatase (STEP) family is expressed within dopaminoceptive neurons of the CNS and is particularly enriched within the basal ganglia and related structures. Alternative splicing produces several isoforms that are found in a number of subcellular compartments, including postsynaptic densities of medium spiny neurons. The variants include STEP(61), a membrane-associated protein, and STEP(46), a cytosolic protein. The C terminals of these two isoforms are identical, whereas the N-terminal domain of STEP(61) contains a novel 172 amino acid sequence that includes several structural motifs not present in STEP(46). Amino acid sequencing revealed a number of potential phosphorylation sites in both STEP isoforms. Therefore, we investigated the role of phosphorylation in regulating STEP activity. Both STEP(61) and STEP(46) are phosphorylated on seryl residues by a cAMP-dependent protein kinase (PKA)-mediated pathway in striatal homogenates. The specific residues phosphorylated in STEP(61) were identified by site-directed mutagenesis and tryptic phosphopeptide mapping as Ser160 and Ser221, whereas the major site of phosphorylation in STEP(46) was shown to be Ser49. Ser160 is located within the unique N terminal of STEP(61). Ser221 and Ser49 are equivalent residues present in STEP(61) and STEP(46), respectively, and are located at the center of the kinase-interacting motif that has been implicated in protein-protein interactions. Phosphorylation at this site decreases the activity of STEP in vitro by reducing its affinity for its substrate. In vivo studies using striatal slices demonstrated that the neurotransmitter dopamine leads to the phosphorylation of STEP via activation of D1 receptors and PKA.
Paul
S
S
The Child Study Center and the Department of Psychiatry, Yale University School of Medicine, New Haven, CT 06520, USA.
Snyder
G L
GL
Yokakura
H
H
Picciotto
M R
MR
Nairn
A C
AC
Lombroso
P J
PJ
D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D020134:Catalytic Domain; D003342:Corpus Striatum / Q000737:chemistry / Q000166:cytology / Q000201:enzymology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004789:Enzyme Activation / Q000502:physiology; D066298:In Vitro Techniques; D008297:Male; D008969:Molecular Sequence Data; D009474:Neurons / Q000737:chemistry / Q000201:enzymology; D010750:Phosphoproteins / Q000032:analysis; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D054558:Protein Tyrosine Phosphatases, Non-Receptor; D051381:Rats; D017207:Rats, Sprague-Dawley; D017447:Receptors, Dopamine D1 / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology
D010750:Phosphoproteins; D010761:Phosphorus Radioisotopes; D017447:Receptors, Dopamine D1; D000255:Adenosine Triphosphate; D017868:Cyclic AMP-Dependent Protein Kinases; D017027:Protein Tyrosine Phosphatases; D054558:Protein Tyrosine Phosphatases, Non-Receptor; C489931:Ptpn5 protein, rat