The dopamine transporter counter-transports potassium to increase the uptake of dopamine

2021 ◽  
Author(s):  
Claus Loland ◽  
Solveig Schmidt ◽  
Mette Malle ◽  
Anne Nielsen ◽  
Søren Bohr ◽  
...  
Keyword(s):  
Ligand Binding ◽  
Dopamine Transporter ◽  
Conformational Dynamics ◽  
Common Mechanism ◽  
Coupled Transport ◽  
Gaba A ◽  
Dopamine Reuptake ◽  
Dopamine Transport ◽  
Neurotransmitter:Sodium Symporter ◽  
The Impact

Abstract The dopamine transporter (DAT) facilitates dopamine reuptake from the extracellular space, and thereby terminates neurotransmission and refills cellular stores of dopamine. DAT belongs to the neurotransmitter:sodium symporter (NSS) family, which includes similar transporters for serotonin, norepinephrine, and GABA. A hallmark of NSS proteins is their ability to utilize the energy stored in the inward-directed Na+ gradient to drive the uphill transport of substrate. Decades ago, it was shown that the serotonin transporter also counter-transports K+, but investigations of K+-coupled transport in other NSSs have been inconclusive. Here, we show that the Drosophila dopamine transporter (dDAT) counter-transports K+. We found that ligand binding to both dDAT and human DAT is inhibited by K+ and that the conformational dynamics of dDAT in K+ is highly divergent from both the apo- and Na+-bound conformations. Furthermore, we found that K+ increased dopamine uptake by purified dDAT reconstituted in liposomes, and we visualized, in real-time, Na+ and K+ fluxes in single proteoliposomes using fluorescent ion indicators. Our results expand on the fundamentals of dopamine transport and prompt a reevaluation of the impact of K+ on other NSSs, including whether K+ counter-transport is a common mechanism for this pharmacologically important protein family.

scholarly journals Probing the impact of temperature and substrates on the conformational dynamics of the Neurotransmitter:Sodium symporter LeuT

2021 ◽  
pp. 167356
Author(s):  
Dionisie Calugareanu ◽  
Ingvar R. Möller ◽  
Solveig G. Schmidt ◽  
Claus J. Loland ◽  
Kasper D. Rand
Keyword(s):  
Conformational Dynamics ◽  
Neurotransmitter:Sodium Symporter ◽  
The Impact

Monitoring the Structural Dynamics of U2 snRNA Via Single-Molecule Förster Resonance Energy Transfer

2018 ◽  
Author(s):  
Alexander Carl DeHaven
Keyword(s):  
Energy Transfer ◽  
Structural Dynamics ◽  
Single Molecule ◽  
Conformational Dynamics ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Förster Resonance Energy Transfer ◽  
U2 Snrna ◽  
Folding Dynamics ◽  
The Impact

This thesis contains four topic areas: a review of single-molecule microscropy methods and splicing, conformational dynamics of stem II of the U2 snRNA, the impact of post-transcriptional modifications on U2 snRNA folding dynamics, and preliminary findings on Mango aptamer folding dynamics.

scholarly journals Predicting pathogenic non-coding SVs disrupting the 3D genome in 1646 whole cancer genomes using multiple instance learning

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Marleen M. Nieboer ◽  
Luan Nguyen ◽  
Jeroen de Ridder
Keyword(s):  
Multiple Instance Learning ◽  
Cancer Diagnostics ◽  
Common Mechanism ◽  
Open Chromatin ◽  
Driver Genes ◽  
3D Genome ◽  
Whole Genomes ◽  
Cancer Genomes ◽  
Cancer Types ◽  
The Impact

AbstractOver the past years, large consortia have been established to fuel the sequencing of whole genomes of many cancer patients. Despite the increased abundance in tools to study the impact of SNVs, non-coding SVs have been largely ignored in these data. Here, we introduce svMIL2, an improved version of our Multiple Instance Learning-based method to study the effect of somatic non-coding SVs disrupting boundaries of TADs and CTCF loops in 1646 cancer genomes. We demonstrate that svMIL2 predicts pathogenic non-coding SVs with an average AUC of 0.86 across 12 cancer types, and identifies non-coding SVs affecting well-known driver genes. The disruption of active (super) enhancers in open chromatin regions appears to be a common mechanism by which non-coding SVs exert their pathogenicity. Finally, our results reveal that the contribution of pathogenic non-coding SVs as opposed to driver SNVs may highly vary between cancers, with notably high numbers of genes being disrupted by pathogenic non-coding SVs in ovarian and pancreatic cancer. Taken together, our machine learning method offers a potent way to prioritize putatively pathogenic non-coding SVs and leverage non-coding SVs to identify driver genes. Moreover, our analysis of 1646 cancer genomes demonstrates the importance of including non-coding SVs in cancer diagnostics.

scholarly journals Dopamine Transporter Genetic Reduction Induces Morpho-Functional Changes in the Enteric Nervous System

Biomedicines ◽  
2021 ◽  
Vol 9 (5) ◽  
pp. 465
Author(s):  
Silvia Cerantola ◽  
Valentina Caputi ◽  
Gabriella Contarini ◽  
Maddalena Mereu ◽  
Antonella Bertazzo ◽  
...  
Keyword(s):  
Nervous System ◽  
Small Intestine ◽  
Enteric Nervous System ◽  
Dopamine Transporter ◽  
Myenteric Plexus ◽  
Receptor Activation ◽  
D1 Receptor ◽  
Functional Changes ◽  
Neurochemical Coding ◽  
The Impact

Antidopaminergic gastrointestinal prokinetics are indeed commonly used to treat gastrointestinal motility disorders, although the precise role of dopaminergic transmission in the gut is still unclear. Since dopamine transporter (DAT) is involved in several brain disorders by modulating extracellular dopamine in the central nervous system, this study evaluated the impact of DAT genetic reduction on the morpho-functional integrity of mouse small intestine enteric nervous system (ENS). In DAT heterozygous (DAT+/−) and wild-type (DAT+/+) mice (14 ± 2 weeks) alterations in small intestinal contractility were evaluated by isometrical assessment of neuromuscular responses to receptor and non-receptor-mediated stimuli. Changes in ENS integrity were studied by real-time PCR and confocal immunofluorescence microscopy in longitudinal muscle-myenteric plexus whole-mount preparations (). DAT genetic reduction resulted in a significant increase in dopamine-mediated effects, primarily via D1 receptor activation, as well as in reduced cholinergic response, sustained by tachykininergic and glutamatergic neurotransmission via NMDA receptors. These functional anomalies were associated to architectural changes in the neurochemical coding and S100β immunoreactivity in small intestine myenteric plexus. Our study provides evidence that genetic-driven DAT defective activity determines anomalies in ENS architecture and neurochemical coding together with ileal dysmotility, highlighting the involvement of dopaminergic system in gut disorders, often associated to neurological conditions.

Conformational dynamics and thermodynamics of protein–ligand binding studied by NMR relaxation

2012 ◽  
Vol 40 (2) ◽  
pp. 419-423 ◽  
Author(s):  
Mikael Akke
Keyword(s):  
Ligand Binding ◽  
Bound States ◽  
Conformational Dynamics ◽  
Nmr Relaxation ◽  
Titration Calorimetry ◽  
Conformational Entropy ◽  
Chain Order ◽  
Ligand Interactions ◽  
Galectin 3 ◽  
Relaxation Experiments

Protein conformational dynamics can be critical for ligand binding in two ways that relate to kinetics and thermodynamics respectively. First, conformational transitions between different substates can control access to the binding site (kinetics). Secondly, differences between free and ligand-bound states in their conformational fluctuations contribute to the entropy of ligand binding (thermodynamics). In the present paper, I focus on the second topic, summarizing our recent results on the role of conformational entropy in ligand binding to Gal3C (the carbohydrate-recognition domain of galectin-3). NMR relaxation experiments provide a unique probe of conformational entropy by characterizing bond-vector fluctuations at atomic resolution. By monitoring differences between the free and ligand-bound states in their backbone and side chain order parameters, we have estimated the contributions from conformational entropy to the free energy of binding. Overall, the conformational entropy of Gal3C increases upon ligand binding, thereby contributing favourably to the binding affinity. Comparisons with the results from isothermal titration calorimetry indicate that the conformational entropy is comparable in magnitude to the enthalpy of binding. Furthermore, there are significant differences in the dynamic response to binding of different ligands, despite the fact that the protein structure is virtually identical in the different protein–ligand complexes. Thus both affinity and specificity of ligand binding to Gal3C appear to depend in part on subtle differences in the conformational fluctuations that reflect the complex interplay between structure, dynamics and ligand interactions.

scholarly journals Conformational entropy of a single peptide controlled under force governs protease recognition and catalysis

2018 ◽  
Vol 115 (45) ◽  
pp. 11525-11530 ◽  
Author(s):  
Marcelo E. Guerin ◽  
Guillaume Stirnemann ◽  
David Giganti
Keyword(s):  
Single Molecule ◽  
Conformational Dynamics ◽  
Conformational Sampling ◽  
Enzymatic Reactions ◽  
Sequence Specificity ◽  
Proteomics Data ◽  
Conformational Entropy ◽  
Substrate Peptide ◽  
The Impact

An immense repertoire of protein chemical modifications catalyzed by enzymes is available as proteomics data. Quantifying the impact of the conformational dynamics of the modified peptide remains challenging to understand the decisive kinetics and amino acid sequence specificity of these enzymatic reactions in vivo, because the target peptide must be disordered to accommodate the specific enzyme-binding site. Here, we were able to control the conformation of a single-molecule peptide chain by applying mechanical force to activate and monitor its specific cleavage by a model protease. We found that the conformational entropy impacts the reaction in two distinct ways. First, the flexibility and accessibility of the substrate peptide greatly increase upon mechanical unfolding. Second, the conformational sampling of the disordered peptide drives the specific recognition, revealing force-dependent reaction kinetics. These results support a mechanism of peptide recognition based on conformational selection from an ensemble that we were able to quantify with a torsional free-energy model. Our approach can be used to predict how entropy affects site-specific modifications of proteins and prompts conformational and mechanical selectivity.

scholarly journals Thermostabilization and purification of the human dopamine transporter (hDAT) in an inhibitor and allosteric ligand bound conformation

2018 ◽  
Author(s):  
Vikas Navratna ◽  
Dilip K. Tosh ◽  
Kenneth A. Jacobson ◽  
Eric Gouaux
Keyword(s):  
Dopamine Transporter ◽  
Limiting Factor ◽  
Molecular Architecture ◽  
Secondary Active Transport ◽  
Reward Seeking ◽  
Dopamine Reuptake ◽  
Seeking Behavior ◽  
Chemical Messenger ◽  
Hormonal Release ◽  
Detergent Micelles

AbstractThe human dopamine transporter(hDAT) plays a major role in dopamine homeostasis and regulation of neurotransmission by clearing dopamine from the extracellular space using secondary active transport. Dopamine is an essential monoamine chemical messenger that regulates reward seeking behavior, motor control, hormonal release, and emotional response in humans. Psychostimulants such as cocaine primarily target the central binding site of hDAT and lock the transporter in an outward-facing conformation, thereby inhibiting dopamine reuptake. The inhibition of dopamine reuptake leads to accumulation of dopamine in the synapse causing heightened signaling. In addition, hDAT is implicated in various neurological disorders and disease-associated neurodegeneration. Despite its significance, the molecular architecture of hDAT and its various conformational states are poorly understood. Instability of hDAT in detergent micelles has been a limiting factor in its successful biochemical, biophysical, and structural characterization. To overcome this hurdle, first we identified ligands that stabilize hDAT in detergent micelles. Then, we screened ∼200 single residue mutants of hDAT using high-throughput scintillation proximity assay, and identified a thermostable variant(I248Y). Here we report a robust strategy to overexpress and successfully purify a thermostable variant of hDAT in an inhibitor and allosteric ligand bound conformation.

scholarly journals Molecular Mechanism of Dopamine Transport by Human Dopamine Transporter

Structure ◽  
2015 ◽  
Vol 23 (11) ◽  
pp. 2171-2181 ◽  
Author(s):  
Mary Hongying Cheng ◽  
Ivet Bahar
Keyword(s):  
Molecular Mechanism ◽  
Dopamine Transporter ◽  
Dopamine Transport

Melatonin receptors limit dopamine reuptake by regulating dopamine transporter cell-surface exposure

2018 ◽  
Vol 75 (23) ◽  
pp. 4357-4370 ◽  
Author(s):  
Abla Benleulmi-Chaachoua ◽  
Alan Hegron ◽  
Marine Le Boulch ◽  
Angeliki Karamitri ◽  
Marta Wierzbicka ◽  
...  
Keyword(s):  
Cell Surface ◽  
Dopamine Transporter ◽  
Melatonin Receptors ◽  
Dopamine Reuptake

Assessment of the mechanical properties of different textile materials used for outer shell of bulletproof vest: woven fabrics for military applications

2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
A.I.H. Fayed ◽  
Y.A. Abo El Amaim ◽  
Ossama Ramy ◽  
Doaa H. Elgohary
Keyword(s):  
Mechanical Properties ◽  
Woven Fabrics ◽  
Outer Shell ◽  
Common Mechanism ◽  
Content Type ◽  
Textile Materials ◽  
Steel Core ◽  
Materials Used ◽  
Bulletproof Vest ◽  
The Impact

Purpose This paper aims to investigate the performance of four different textile materials used as an outer shell of the bulletproof vest. Design/methodology/approach In this paper, four different textile materials were used, polyurethane treatment was applied as a surface coating for the woven samples. Mechanical properties were conducted for all samples; scanning electron microscope and X-ray energy disperse spectroscopy were executed to show the surface morphology of samples and the chemical composition of the coating material. Findings One-way ANOVA was used to statistically analyse the results, which proved that all variables were highly significantly affected by using different textile materials, despite the stiffness variable being not significantly affected by textile materials. An overall evaluation was done using radar chart, demonstrated that Cordura material accomplished the best functional performance, using two types of calibres 7.62 × 54 mild steel core and 7.62 × 54 armour piercing incendiary; the common mechanism was localized burn because of the incendiary effect of the projectile in addition to tearing mechanism starting from inside because of penetration effect of the steel core. Originality/value This work was addressed to analyse the impact of using four different materials on its performance as the outer shell of bulletproof vest to achieve the desired degree of protection.

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