Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.4.1 (phosphodiesterase)
 18,767 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Several transmembrane transporters of organic compounds are regulated by phosphorylation/dephosphorylation mechanisms. The aim of this study was to investigate the possible regulation of the human extraneuronal monoamine transporter, hEMT, by these mechanisms. The experiments were performed using HEK293 cells stably transfected with pcDNA3hEMT (293hEMT). The characteristics of hEMT-mediated uptake of [3H]1-methyl-4-phenylpyridinium ([3H]MPP+) were studied by incubating the cells at 37 degrees C for 1 min with 200 nM [3H]MPP+. Uptake of [3H]MPP+ by 293hEMT cells was not affected or only slightly reduced by modulators of protein kinase A, protein kinase C, or protein kinase G. It was not affected by an inhibitor of protein tyrosine kinase and was reduced by mitogen-activated protein kinase inhibitors. Uptake of [3H]MPP+ by 293hEMT cells was independent of extracellular Ca2+ and strongly reduced by Ca2+/calmodulin pathway inhibitors. Uptake of [3H]MPP+ by 293hEMT cells was strongly reduced in the presence of non-selective phosphodiesterase inhibitors (IBMX, caffeine, theophylline). The effect of IBMX was independent of extracellular Ca2+ its IC50 was found to be 82.0 microM (66.2-101.6 microM; n=4), and its inhibitory effect resulted from a significant decrease in the maximal velocity of [3H]MPP+ uptake, with no change in the Michaelis-Menten constant. [3H]MPP+ uptake was reduced by 8-methoxy-methyl-IBMX, a selective inhibitor of the Ca2+/calmodulin-dependent phosphodiesterase (PDE1), but not by zaprinast, a selective inhibitor of PDE5. Uptake of [3H]MPP+ by 293hEMT cells was strongly reduced by protein tyrosine phosphatase inhibitors, by an alkaline phosphatase inhibitor and, by contrast. showed an increase in the presence of exogenous alkaline phosphatase. In conclusion, these results suggest that hEMT is regulated by phosphorylation/dephosphorylation mechanisms, being active in the dephosphorylated state.
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PMID:Regulation of human extraneuronal monoamine transporter (hEMT) expressed in HEK293 cells by intracellular second messenger systems. 1177 2

Several transmembrane transporters of organic compounds are regulated by phosphorylation/dephosphorylation mechanisms. The aim of this study was to investigate the possible regulation of the intestinal uptake of organic cations by these mechanisms. The intestinal apical uptake of 1-methyl-4-phenylpyridinium (MPP(+)) was studied by incubating Caco-2 cells at 37 degrees for 5 min with 200 nM (3)H-MPP(+). Uptake of (3)H-MPP(+) by Caco-2 cells was not affected by activators of protein kinase G, and was not affected or slightly reduced (by 15-20%) by activators of protein kinase A or protein kinase C. Uptake of (3)H-MPP(+) by Caco-2 cells was reduced in a concentration-dependent manner by non-selective phosphodiesterase inhibitors (3-isobutyl-1-methylxanthine (IBMX), caffeine, teophylline). The IC(50) of IBMX was found to be 119 microM (102-138; n=9). Uptake of (3)H-MPP(+) by Caco-2 cells was not affected by inhibition of protein tyrosine kinase, but it was concentration-dependently reduced in the presence of inhibitors of mitogen-activated protein kinase. Uptake of (3)H-MPP(+) by Caco-2 cells was strongly reduced by Ca(2+)/calmodulin-mediated pathway inhibitors, but it was not dependent on extracellular Ca(2+). Our results suggest that the intestinal apical uptake of MPP(+) is regulated by phosphorylation/dephosphorylation mechanisms, being most probably active in the dephosphorylated state. Moreover, uptake of (3)H-MPP(+) by Caco-2 cells and by the extraneuronal monoamine transporter (EMT) are regulated in a very similar manner, suggesting an important participation of EMT in the intestinal uptake of this compound.
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PMID:Uptake of (3)H-1-methyl-4-phenylpyridinium ((3)H-MPP(+)) by human intestinal Caco-2 cells is regulated by phosphorylation/dephosphorylation mechanisms. 1199 99