Other search tools

About this data

The publication data currently available has been vetted by Vanderbilt faculty, staff, administrators and trainees. The data itself is retrieved directly from NCBI's PubMed and is automatically updated on a weekly basis to ensure accuracy and completeness.

If you have any questions or comments, please contact us.

Results: 21 to 29 of 29

Publication Record


Inhibitors of the serotonin transporter protein (SERT): the design and synthesis of biotinylated derivatives of 3-(1,2,3,6-tetrahydro-pyridin-4-yl)-1H-indoles. High-affinity serotonergic ligands for conjugation with quantum dots.
Tomlinson ID, Mason JN, Blakely RD, Rosenthal SJ
(2005) Bioorg Med Chem Lett 15: 5307-10
MeSH Terms: Biotinylation, Humans, Imidazoles, Indoles, Ligands, Molecular Structure, Quantum Dots, Serotonin Plasma Membrane Transport Proteins, Serotonin Uptake Inhibitors
Show Abstract · Added July 10, 2013
There is a growing demand for compounds with specificity for the serotonin transporter protein (SERT) that can be conjugated to cadmium selenide/zinc sulfide core shell nanocrystals. This letter describes the design and synthesis of two different biotinylated SERT antagonists that can be attached to streptavidin-coated cadmium selenide/zinc sulfide core shell nanocrystals.
1 Communities
2 Members
0 Resources
9 MeSH Terms
Preferential Interaction between the dopamine D2 receptor and Arrestin2 in neostriatal neurons.
Macey TA, Gurevich VV, Neve KA
(2004) Mol Pharmacol 66: 1635-42
MeSH Terms: Animals, Arrestins, Biotinylation, Cell Line, Cells, Cultured, Male, Neostriatum, Neurons, Phosphoproteins, Protein Transport, Rats, Rats, Sprague-Dawley, Receptors, Dopamine D2, Recombinant Proteins, Transfection
Show Abstract · Added December 10, 2013
Dopamine D2 receptor interactions with arrestins and arrestin-dependent internalization have been characterized using heterologously expressed D2 receptor and arrestins. The purpose of this study was to investigate D2 receptor interaction with endogenous arrestins. Arrestin2 and arrestin3 in striatal homogenates bound to the third cytoplasmic loop of the D2 receptor, and purified arrestin2 and arrestin3 bound to the second and third loops and C terminus of the D2 receptor, in a glutathione S-transferase pull-down assay. In NS20Y neuroblastoma cells expressing an enhanced green-fluorescent protein-tagged D2 receptor (D2-EGFP), 2-h D2 agonist stimulation enhanced the colocalization of D2-EGFP with endogenous arrestin2 and arrestin3. These results suggest that the D2 receptor has the intrinsic ability to bind both nonvisual arrestins. Agonist treatment of D2-EGFP NS20Y cells induced D2 receptor internalization (36-46%) that was maximal within 20 min, but that was prevented by small interfering RNA-induced depletion of arrestin2 and arrestin3. In neostriatal neurons, 2-h agonist treatment selectively increased the colocalization of the endogenous D2 receptor with arrestin2, whereas receptor colocalization with arrestin3 was reduced. Agonist stimulation caused translocation of arrestin2, but not arrestin3, to the membrane in neurons and selectively enhanced the coimmunoprecipitation of the D2 receptor and arrestin2. All three measures of receptor/arrestin interaction (colocalization, translocation, and coprecipitation) demonstrated selective agonist-induced interaction between the D2 receptor and arrestin2 in neurons.
0 Communities
1 Members
0 Resources
15 MeSH Terms
Reverse endocytosis of transmembrane ephrin-B ligands via a clathrin-mediated pathway.
Parker M, Roberts R, Enriquez M, Zhao X, Takahashi T, Pat Cerretti D, Daniel T, Chen J
(2004) Biochem Biophys Res Commun 323: 17-23
MeSH Terms: Animals, Biotinylation, Blotting, Western, CHO Cells, Cell Membrane, Cell Separation, Cells, Cultured, Clathrin, Cricetinae, DNA, Complementary, Dynamins, Endocytosis, Ephrin-B1, Flow Cytometry, Genes, Dominant, Genetic Vectors, Humans, Ligands, Microscopy, Confocal, Microscopy, Electron, Mutation, Potassium, Receptors, Eph Family, Signal Transduction, Time Factors, Transfection, Umbilical Veins
Show Abstract · Added August 27, 2013
Eph/ephrin receptors and ligands mediate cell-cell interaction through reciprocal signaling upon juxtacrine contact, and play a critical role in embryonic patterning, neuronal targeting, and vascular assembly. To study transmembrane ephrin-B ligand trafficking, we determined the cellular localization of ephrin-B1-GFP upon engagement by EphB1. Under normal culture conditions ephrin-B1-GFP is localized to the plasma membrane, mostly at the lateral cell borders. Addition of soluble EphB1-Fc receptor induces ephrin-B1-GFP clustering on the cell surface and subsequent internalization, as judged by biochemical studies, electron microscopy, and co-localization with endosomal markers. A dominant-negative mutant of dynamin or potassium depletion blocks ephrin-B1 endocytosis. These results suggest that ephrin-B1 internalization is an active receptor-mediated process that utilizes the clathrin-mediated endocytic pathway.
Copyright 2004 Elsevier Inc.
1 Communities
2 Members
0 Resources
27 MeSH Terms
N-terminal phosphorylation of the dopamine transporter is required for amphetamine-induced efflux.
Khoshbouei H, Sen N, Guptaroy B, Johnson L', Lund D, Gnegy ME, Galli A, Javitch JA
(2004) PLoS Biol 2: E78
MeSH Terms: Amphetamines, Aspartic Acid, Biotinylation, Cell Line, Cell Membrane, Cells, Cultured, Dopamine Plasma Membrane Transport Proteins, Electrophysiology, Humans, Immunoblotting, Kinetics, Molecular Sequence Data, Mutation, Perfusion, Phenotype, Phosphorylation, Plasmids, Protein Kinase C, Protein Structure, Tertiary, Serine, Transfection, Tyramine
Show Abstract · Added December 10, 2013
Amphetamine (AMPH) elicits its behavioral effects by acting on the dopamine (DA) transporter (DAT) to induce DA efflux into the synaptic cleft. We previously demonstrated that a human DAT construct in which the first 22 amino acids were truncated was not phosphorylated by activation of protein kinase C, in contrast to wild-type (WT) DAT, which was phosphorylated. Nonetheless, in all functions tested to date, which include uptake, inhibitor binding, oligomerization, and redistribution away from the cell surface in response to protein kinase C activation, the truncated DAT was indistinguishable from the full-length WT DAT. Here, however, we show that in HEK-293 cells stably expressing an N-terminal-truncated DAT (del-22 DAT), AMPH-induced DA efflux is reduced by approximately 80%, whether measured by superfusion of a population of cells or by amperometry combined with the patch-clamp technique in the whole cell configuration. We further demonstrate in a full-length DAT construct that simultaneous mutation of the five N-terminal serine residues to alanine (S/A) produces the same phenotype as del-22-normal uptake but dramatically impaired efflux. In contrast, simultaneous mutation of these same five serines to aspartate (S/D) to simulate phosphorylation results in normal AMPH-induced DA efflux and uptake. In the S/A background, the single mutation to Asp of residue 7 or residue 12 restored a significant fraction of WT efflux, whereas mutation to Asp of residues 2, 4, or 13 was without significant effect on efflux. We propose that phosphorylation of one or more serines in the N-terminus of human DAT, most likely Ser7 or Ser12, is essential for AMPH-induced DAT-mediated DA efflux. Quite surprisingly, N-terminal phosphorylation shifts DAT from a "reluctant" state to a "willing" state for AMPH-induced DA efflux, without affecting inward transport. These data raise the therapeutic possibility of interfering selectively with AMPH-induced DA efflux without altering physiological DA uptake.
0 Communities
1 Members
0 Resources
22 MeSH Terms
Calmodulin interacts with the third intracellular loop of the serotonin 5-hydroxytryptamine1A receptor at two distinct sites: putative role in receptor phosphorylation by protein kinase C.
Turner JH, Gelasco AK, Raymond JR
(2004) J Biol Chem 279: 17027-37
MeSH Terms: Amino Acid Motifs, Amino Acid Sequence, Animals, Binding Sites, Biotinylation, Brain, CHO Cells, Calmodulin, Cattle, Cricetinae, Dose-Response Relationship, Drug, Fibroblasts, Fluorescence Resonance Energy Transfer, Fluorometry, Kinetics, Molecular Sequence Data, Peptides, Phosphorylation, Plasmids, Precipitin Tests, Protein Binding, Protein Conformation, Protein Kinase C, Protein Structure, Secondary, Protein Structure, Tertiary, Rats, Receptor, Serotonin, 5-HT1A, Sequence Homology, Amino Acid, Signal Transduction, Spectrometry, Fluorescence, Surface Plasmon Resonance, Time Factors, Transfection
Show Abstract · Added July 23, 2020
The serotonin 5-HT(1A) receptor couples to heterotrimeric G proteins and intracellular second messengers, yet no studies have investigated the possible role of additional receptor-interacting proteins in 5-HT(1A) receptor signaling. We have found that the ubiquitous Ca(2+)-sensor calmodulin (CaM) co-immunoprecipitates with the 5-HT(1A) receptor in Chinese hamster ovary fibroblasts. The human 5-HT(1A) receptor contains two putative CaM binding motifs, located in the N- and C-terminal juxtamembrane regions of the third intracellular loop of the receptor. Peptides encompassing both the N-terminal (i3N) and C-terminal (i3C) CaM-binding domains were tested for CaM binding. Using in vitro binding assays in combination with gel shift analysis, we demonstrated Ca(2+)-dependent formation of complexes between CaM and both peptides. We determined kinetic data using a combination of BIAcore surface plasmon resonance (SPR) and dansyl-CaM fluorescence. SPR analysis gave an apparent K(D) of approximately 110 nm for the i3N peptide and approximately 700 nm for the i3C peptide. Both peptides also caused characteristic shifts in the fluorescence emission spectrum of dansyl-CaM, with apparent affinities of 87 +/- 23 nm and 1.70 +/- 0.16 microm. We used bioluminescence resonance energy transfer to show that CaM interacts with the 5-HT(1A) receptor in living cells, representing the first in vivo evidence of a G protein-coupled receptor interacting with CaM. Finally, we showed that CaM binding and phosphorylation of the 5-HT(1A) receptor i3 loop peptides by protein kinase C are antagonistic in vitro, suggesting a possible role for CaM in the regulation of 5-HT(1A) receptor phosphorylation and desensitization. These data suggest that the 5-HT(1A) receptor contains high and moderate affinity CaM binding regions that may play important roles in receptor signaling and function.
0 Communities
1 Members
0 Resources
MeSH Terms
Ethnicity-dependent polymorphism in Na+-taurocholate cotransporting polypeptide (SLC10A1) reveals a domain critical for bile acid substrate recognition.
Ho RH, Leake BF, Roberts RL, Lee W, Kim RB
(2004) J Biol Chem 279: 7213-22
MeSH Terms: African Continental Ancestry Group, Alleles, Amino Acid Sequence, Asian Continental Ancestry Group, Bile Acids and Salts, Biological Transport, Biotinylation, Carrier Proteins, Cell Line, Cell Membrane, Dose-Response Relationship, Drug, Electrophoresis, Polyacrylamide Gel, European Continental Ancestry Group, Gene Frequency, Genotype, Glycosylation, HeLa Cells, Hepatocytes, Hispanic Americans, Humans, Microscopy, Confocal, Molecular Sequence Data, Organic Anion Transporters, Sodium-Dependent, Plasmids, Polymorphism, Genetic, Polymorphism, Single Nucleotide, Protein Binding, Protein Structure, Tertiary, Sequence Homology, Amino Acid, Substrate Specificity, Symporters, Taurocholic Acid, Vaccinia virus
Show Abstract · Added March 5, 2014
The key transporter responsible for hepatic uptake of bile acids from portal circulation is Na+-taurocholate cotransporting polypeptide (NTCP, SLC10A1). This transporter is thought to be critical for the maintenance of enterohepatic recirculation of bile acids and hepatocyte function. Therefore, functionally relevant polymorphisms in this transporter would be predicted to have an important impact on bile acid homeostasis/liver function. However, little is known regarding genetic heterogeneity in NTCP. In this study, we demonstrate the presence of multiple single nucleotide polymorphisms in NTCP in populations of European, African, Chinese, and Hispanic Americans. Specifically four nonsynonymous single nucleotide polymorphisms associated with a significant loss of transport function were identified. Cell surface biotinylation experiments indicated that the altered transport activity of T668C (Ile223-->Thr), a variant seen only in African Americans, was due at least in part to decreased plasma membrane expression. Similar expression patterns were observed when the variant alleles were expressed in HepG2 cells, and plasma membrane expression was assessed using immunofluorescence confocal microscopy. Interestingly the C800T (Ser267-->Phe) variant, seen only in Chinese Americans, exhibited a near complete loss of function for bile acid uptake yet fully normal transport function for the non-bile acid substrate estrone sulfate, suggesting this position may be part of a region in the transporter critical and specific for bile acid substrate recognition. Accordingly, our study indicates functionally important polymorphisms in NTCP exist and that the likelihood of being carriers of such polymorphisms is dependent on ethnicity.
2 Communities
2 Members
2 Resources
33 MeSH Terms
Membrane topology of Bves/Pop1A, a cell adhesion molecule that displays dynamic changes in cellular distribution during development.
Knight RF, Bader DM, Backstrom JR
(2003) J Biol Chem 278: 32872-9
MeSH Terms: 3T3 Cells, Amino Acid Sequence, Animals, Asparagine, Avian Proteins, Biotinylation, COS Cells, Cell Adhesion, Cell Adhesion Molecules, Cell Line, Cell Membrane, Chick Embryo, Cytoplasm, Detergents, Electrophoresis, Polyacrylamide Gel, Endoplasmic Reticulum, Gene Deletion, Gene Expression Regulation, Developmental, Glycosylation, Immunohistochemistry, Mice, Models, Biological, Models, Genetic, Molecular Sequence Data, Muscle Proteins, Myocardium, Precipitin Tests, Protein Structure, Tertiary, Rats, Sequence Homology, Amino Acid, Tissue Distribution, Transfection
Show Abstract · Added September 28, 2015
We investigated the membrane topology of Bves/Pop1A as a foundation to dissect the molecular basis and function of Bves/Pop1A trafficking during development. Bves contains two asparagine-linked glycosylation sites within the amino terminus and three putative membrane domains. Therefore, glycosylation assays were performed to determine if the amino terminus of Bves is delivered into the endoplasmic reticulum lumen and glycosylated. We establish that Bves from chick heart and transfected cells is glycosylated, implying that the amino terminus of cell surface molecules is extracellular. Three biochemically distinct approaches were utilized to determine the orientation of the carboxyl terminus of Bves. First, glycosylation of Bves at exogenous sites within the carboxyl terminus was only observed in a construct that lacked the third membrane domain, which presumably reversed the orientation of the carboxyl terminus. Second, co-expression of full-length Bves with soluble, carboxyl-terminal Bves constructs that reside in different subcellular compartments revealed that Bves-Bves interactions occur in the cytoplasm. Third, the immunoreactivity of endogenous Bves at the cell surface of epicardial cells was dramatically enhanced with detergent. These results suggest that the membrane topology of cell surface Bves/Pop1A is composed of an extracellular amino terminus, three transmembrane domains, and a cytoplasmic carboxyl terminus. We therefore hypothesize that the carboxyl terminus regulates the cellular distribution of Bves/Pop1A during coronary vessel development.
1 Communities
1 Members
0 Resources
32 MeSH Terms
Inhibition of pRb phosphorylation and cell cycle progression by an antennapedia-p16(INK4A) fusion peptide in pancreatic cancer cells.
Fujimoto K, Hosotani R, Miyamoto Y, Doi R, Koshiba T, Otaka A, Fujii N, Beauchamp RD, Imamura M
(2000) Cancer Lett 159: 151-8
MeSH Terms: Amino Acid Sequence, Antennapedia Homeodomain Protein, Biotinylation, Carrier Proteins, Cell Cycle, Cell Division, Cyclin A, Cyclin-Dependent Kinase Inhibitor p16, Dose-Response Relationship, Drug, Homeodomain Proteins, Humans, Molecular Sequence Data, Nuclear Proteins, Pancreatic Neoplasms, Phosphorylation, Recombinant Fusion Proteins, Retinoblastoma Protein, Transcription Factors, Tumor Cells, Cultured
Show Abstract · Added June 14, 2013
In this study, we examined whether or not a small peptide derived from p16(INK4A) protein with the antennapedia carrier sequence could inhibit the growth of pancreatic cancer cells through the inhibition of cell cycle progression. Growth inhibition by the p16-derived peptide was observed in a time- and dose-dependent manner in AsPC-1 and BxPC-3 cells (p16-negative and pRb-positive), whereas Saos-2 cells (p16-positive and pRb-negative) showed no inhibitory effect. In AsPC-1 and BxPC-3 cells, the proportion of cells in the G(1) phase markedly increased 48 h after treatment with 20 microM p16-derived peptide. Cell-cycle analysis of Saos-2 cells showed little change during the entire period of treatment. Immunoblot analysis showed inhibition of pRb phosphorylation after treatment of BxPC-3 with 10 microM p16 peptide. Furthermore, the p16 peptide caused a decrease in cyclin A at later times of treatment. These results demonstrate that the p16-derived peptide can inhibit the growth of p16-negative and pRb-positive pancreatic cancer cells by means of G(1) phase cell cycle arrest resulting from the inhibition of pRb phosphorylation. Restoration of p16/pRb tumor-suppressive pathway by re-expression of p16(INK4A) may play a therapeutic role in the treatment of pancreatic cancer.
1 Communities
1 Members
0 Resources
19 MeSH Terms
Phosphorylation and sequestration of serotonin transporters differentially modulated by psychostimulants.
Ramamoorthy S, Blakely RD
(1999) Science 285: 763-6
MeSH Terms: Antidepressive Agents, Biogenic Monoamines, Biotinylation, Carrier Proteins, Cell Line, Central Nervous System Agents, Cocaine, Dextroamphetamine, Enzyme Activation, Humans, Ligands, Membrane Glycoproteins, Membrane Transport Proteins, Models, Biological, Nerve Tissue Proteins, Neurotransmitter Agents, Phosphorylation, Protein Kinase C, Protein Kinases, Serotonin, Serotonin Antagonists, Serotonin Plasma Membrane Transport Proteins, Serotonin Uptake Inhibitors, Tetradecanoylphorbol Acetate
Show Abstract · Added July 10, 2013
Many psychotropic drugs interfere with the reuptake of dopamine, norepinephrine, and serotonin. Transport capacity is regulated by kinase-linked pathways, particularly those involving protein kinase C (PKC), resulting in transporter phosphorylation and sequestration. Phosphorylation and sequestration of the serotonin transporter (SERT) were substantially impacted by ligand occupancy. Ligands that can permeate the transporter, such as serotonin or the amphetamines, prevented PKC-dependent SERT phosphorylation. Nontransported SERT antagonists such as cocaine and antidepressants were permissive for SERT phosphorylation but blocked serotonin effects. PKC-dependent SERT sequestration was also blocked by serotonin. These findings reveal activity-dependent modulation of neurotransmitter reuptake and identify previously unknown consequences of amphetamine, cocaine, and antidepressant action.
1 Communities
1 Members
0 Resources
24 MeSH Terms