Identification of a cyclic adenosine 3′,5′-monophosphate-dependent protein kinase phosphorylation site in the carboxy terminal tail of human D1 dopamine receptor

Zamanillo, Daniel; Casanova, Emilio; Alonso‐Llamazares, Ana; Ovalle, Sergio; Chinchetru, Miguel A.; Calvo, Pedro

doi:10.1016/0304-3940(95)11428-y

Cited by 20 publications

(11 citation statements)

References 17 publications

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“…Like most G-protein-coupled receptors, D1 receptor undergoes both a second messenger-dependent kinase and G-protein-coupled receptor kinase (GRK)-mediated phosphorylation reaction that leads to its desensitization (42,43). Although in the present study, we found increased PKC activity, it is reported that PKC cannot the phosphorylate D1A receptor because the receptor lacks the phosphorylation sites for PKC (44). Because GRKs are involved in the phosphorylation of D1 receptors, resulting in their desensitization, it is of interest that GRK-2 and -4 activities and expression are increased in patients with essential hypertension (13,45).…”

Section: Discussioncontrasting

confidence: 53%

Tempol Reduces Oxidative Stress, Improves Insulin Sensitivity, Decreases Renal Dopamine D1 Receptor Hyperphosphorylation, and Restores D1 Receptor–G-Protein Coupling and Function in Obese Zucker Rats

et al. 2005

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Oxidative stress plays a pathogenic role in hypertension, particularly the one associated with diabetes and obesity. Here, we test the hypothesis that renal dopamine D1 receptor dysfunction in obese Zucker rats is caused by oxidative stress. One group each from lean and obese Zucker rats received tempol, a superoxide dismutase mimetic in drinking water for 2 weeks. Obese animals were hypertensive, hyperglycemic, and hyperinsulinemic, exhibited renal oxidative stress, and increased protein kinase C activity. Also, there was hyperphosphorylation of D1 receptor, defective receptor-G-protein coupling, blunted dopamine-induced Na ؉ -K ؉ -ATPase inhibition, and diminished natriuretic response to D1 receptor agonist, SKF-38393. However, obese animals had elevated levels of plasma nitric oxide and urinary cGMP. In addition, L-N-nitroarginine and sodium nitroprusside showed similar effect on blood pressure in lean and obese rats. In obese animals, tempol reduced blood pressure, blood glucose, insulin, renal oxidative stress, and protein kinase C activity. Tempol also decreased D1 receptor phosphorylation and restored receptor G-protein coupling. Dopamine inhibited Na ؉ -K ؉ -ATPase activity, and SKF-38393 elicited a natriuretic response in tempol-treated obese rats. Thus in obese Zucker rats, tempol ameliorates oxidative stress and improves insulin sensitivity. Consequently, hyperphosphorylation of D1 receptor is reduced, leading to restoration of receptor-G-protein coupling and the natriuretic response to SKF-38393. Diabetes 54: 2219 -2226, 2005

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Section: Discussioncontrasting

confidence: 53%

Tempol Reduces Oxidative Stress, Improves Insulin Sensitivity, Decreases Renal Dopamine D1 Receptor Hyperphosphorylation, and Restores D1 Receptor–G-Protein Coupling and Function in Obese Zucker Rats

et al. 2005

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“…The primate D 1 receptor has three potential sites of PKA-catalyzed phosphorylation in cytoplasmic domains. Although the predicted rank order of preference of the sites for phosphorylation by PKA is Thr136 Ͼ Thr268 ϭ Ser380 (Kennelly and Krebs, 1991), we mutated the two residues suggested by previous work to be involved in desensitization of the D 1 receptor (Thr268; Jiang and Sibley, 1999), or to be a site of PKA-catalyzed phosphorylation (Ser380; Zamanillo et al, 1995). Whereas mutation of Ser380 had no effect on dopamine-stimulated phosphorylation of the receptor, mutation of Thr268 prevented the dopamine-induced incorporation of phosphate.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, desensitization of the D 1 receptor is blunted in cells deficient in PKA (Ventura and Sibley, 2000), and mutation of a potential site of PKA phosphorylation of the D 1 receptor, Thr268, reduces the rate of agonist-induced desensitization (Jiang and Sibley, 1999). Ser380 has also been proposed to be a site of phosphorylation by PKA, because a peptide comprised of D 1 receptor amino acid residues 372 to 442 is phosphorylated by PKA on Ser380 (Zamanillo et al, 1995).…”

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confidence: 99%

Regulation of Dopamine D₁Receptor Trafficking by Protein Kinase A-Dependent Phosphorylation

2002

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The aim of this study was to use pharmacological inhibition of protein kinase A and mutation of potential protein kinase A phosphorylation sites to determine the role of protein kinase A-catalyzed phosphorylation of the dopamine D 1 receptor in agonist-stimulated desensitization and internalization of the receptor. To facilitate purification and imaging of the D 1 receptor, we attached a polyhistidine tag to the amino terminus and enhanced green fluorescent protein to the carboxyl terminus of the receptor (D 1 -EGFP). D 1 -EGFP was similar to the untagged D 1 receptor in terms of affinity for agonist and antagonist ligands, coupling to G proteins, and stimulation of cyclic AMP accumulation. D 1 -EGFP and two mutants in which either Thr268 or Ser380 was replaced with Ala were stably expressed in NS20Y neuroblastoma cells. Pretreatment with the protein kinase A inhibitor H-89 (N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide) or substitution of Ala for Thr268 reduced agonist-stimulated phosphorylation of the receptor and resulted in diminished trafficking of the receptor to the perinuclear region of the cell. Substitution of Ala for Thr268 had no effect, however, on agonist-induced receptor sequestration or desensitization of cyclic AMP accumulation. Substitution of Ala for Ser380 had no effect on D 1 receptor phosphorylation, sequestration, desensitization, or trafficking to the perinuclear region. We conclude that protein kinase A-dependent phosphorylation of the D 1 receptor on Thr268 regulates a late step in the sorting of the receptor to the perinuclear region of the cell, but that phosphorylation of Thr268 is not required for receptor sequestration or maximal desensitization of cyclic AMP accumulation.

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“…Also depicted are hDAT-chimeric constructs in which hDAT tail sequence was substituted by amino acid residues encoding the COOH-terminal tail of dopamine D1 (hDAT-D1) or D5 (hDAT-D5) receptors. Although hDAT-tr1 and the hDAT-D5 mutant lost all putative COOH-terminal phosphorylation sites, hDAT-tr2 retained consensus sequence for protein kinase C sites while the hDAT-D1 mutant contains only one functional protein kinase A site (85). mutants are listed in Table I.…”

Section: Resultsmentioning

confidence: 99%

The Dopamine Transporter Carboxyl-terminal Tail

Lee

Pristupa

Ciliax

et al. 1996

Journal of Biological Chemistry

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In order to delineate structural motifs regulating substrate affinity and recognition for the human dopamine transporter (DAT), we assessed [ The Na ϩ -and Cl Ϫ -dependent nigral-striatal presynaptic dopamine transporter (DAT), 1 by mediating the reuptake of dopamine into the cell, significantly regulates the availability of synaptic dopamine (DA) to effectively interact with multiple pre-and postsynaptic dopamine receptors (for reviews see Refs. 1-4). The DAT is the presumed major target site for the accumulation of the dopaminergic neurotoxin, 1-methyl-phenylpyridine (MPP ϩ ), producing Parkinson's like symptoms (5), and the initial site of action of psychostimulants and drugs of abuse, such as amphetamine, methylphenidate, and cocaine to elicit psychomotor behavior associated with euphoria, self-reward, and addiction (see . The isolation of cDNA clones for the mouse (11), rat (12-14), bovine (15) and human has revealed that it is a member of a large gene family of Na ϩ -and Cl Ϫ -dependent transporters (20 -22) with particular homology to neurotransmitter transport proteins for noradrenaline (NE) (23) and serotonin (24). As with other members of the family, hydropathic analysis and molecular modeling (25) of the amino acid sequence of the DAT predicts a common topology of 12 putative transmembrane (TM) domains, a large extracellular loop between TM 3 and 4 containing numerous consensus sequences for N-linked glycosylation, and potential sites for phosphorylation by protein kinase A, protein kinase C, and CaM kinase II within putative intracellular domains and in the amino and carboxyl termini. The human and rat DATs are encoded by polypeptides of ϳ69 kDa with 92% overall amino acid sequence homology. Upon heterologous expression in mammalian cells, the cloned DAT confers Na ϩ -and Cl Ϫ -dependent bidirectional dopamine transport (18, 26) and cytotoxicity to MPP ϩ (27, 28), which is sensitive to the inhibition by DAT antagonists including cocaine, methylpheni-

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Identification of a cyclic adenosine 3′,5′-monophosphate-dependent protein kinase phosphorylation site in the carboxy terminal tail of human D1 dopamine receptor

Cited by 20 publications

References 17 publications

Tempol Reduces Oxidative Stress, Improves Insulin Sensitivity, Decreases Renal Dopamine D1 Receptor Hyperphosphorylation, and Restores D1 Receptor–G-Protein Coupling and Function in Obese Zucker Rats

Tempol Reduces Oxidative Stress, Improves Insulin Sensitivity, Decreases Renal Dopamine D1 Receptor Hyperphosphorylation, and Restores D1 Receptor–G-Protein Coupling and Function in Obese Zucker Rats

Regulation of Dopamine D₁Receptor Trafficking by Protein Kinase A-Dependent Phosphorylation

The Dopamine Transporter Carboxyl-terminal Tail

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Identification of a cyclic adenosine 3′,5′-monophosphate-dependent protein kinase phosphorylation site in the carboxy terminal tail of human D1 dopamine receptor

Cited by 20 publications

References 17 publications

Tempol Reduces Oxidative Stress, Improves Insulin Sensitivity, Decreases Renal Dopamine D1 Receptor Hyperphosphorylation, and Restores D1 Receptor–G-Protein Coupling and Function in Obese Zucker Rats

Tempol Reduces Oxidative Stress, Improves Insulin Sensitivity, Decreases Renal Dopamine D1 Receptor Hyperphosphorylation, and Restores D1 Receptor–G-Protein Coupling and Function in Obese Zucker Rats

Regulation of Dopamine D1Receptor Trafficking by Protein Kinase A-Dependent Phosphorylation

The Dopamine Transporter Carboxyl-terminal Tail

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Regulation of Dopamine D₁Receptor Trafficking by Protein Kinase A-Dependent Phosphorylation