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Find video protocols related to scientific articles indexed in Pubmed.
Selective Inhibition of Mutant Isocitrate Dehydrogenase 1 (IDH1) via Disruption of a Metal Binding Network by an Allosteric Small Molecule.
J. Biol. Chem.
PUBLISHED: 11-14-2014
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Cancer-associated point mutations in isocitrate dehydrogenase 1 and 2 (IDH1, IDH2) confer a neomorphic enzymatic activity: the reduction of alpha-ketoglutarate (?KG) to D-2-hydroxyglutaric acid (2HG), which is proposed to act as an oncogenic metabolite by inducing hypermethylation of histones and DNA. While selective inhibitors of mutant IDH1 and IDH2 have been identified and are currently under investigation as potential cancer therapeutics, the mechanistic basis for their selectivity is not yet well-understood. A high-throughput screen for selective inhibitors of IDH1 bearing the oncogenic mutation R132H identified Compound 1, a bis-imidazole phenol that inhibits 2HG production in cells. We investigated the mode of inhibition of Compound 1 and a previously published IDH1 mutant inhibitor with a different chemical scaffold. Steady-state kinetics and biophysical studies show that both of these compounds selectively inhibit mutant IDH1 by binding to an allosteric site, and that inhibition is competitive with respect to Mg(2+). A crystal structure of Compound 1 complexed with R132H IDH1 indicates that the inhibitor binds at the dimer interface and makes a direct contact with a residue involved in binding of the catalytically essential divalent cation. These results show that targeting a divalent cation binding residue can enable selective inhibition of mutant IDH1, and suggest that differences in magnesium binding between wild-type and mutant enzymes may contribute to the inhibitors' selectivity for the mutant enzyme.
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Advances in understanding the mechanism and improved stability of the synthesis of ammonia from air and water in hydroxide suspensions of nanoscale Fe?O?.
Inorg Chem
PUBLISHED: 09-23-2014
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We report a mechanism of electrochemical ammonia (NH3) production via an iron intermediate in which H2 and NH3 are cogenerated by different electron-transfer pathways. Solar thermal can contribute to the energy to drive this synthesis, resulting in a STEP, solar thermal electrochemical process, for NH3. Enhancements are presented to this carbon dioxide (CO2)-free synthesis, which uses suspensions of nano-Fe2O3 in high-temperature hydroxide electrolytes at nickel and Monel electrodes. In a 200 °C molten eutectic Na(0.5)K(0.5)OH electrolyte, the 3 Faraday efficiency per mole of synthesized NH3, ?(NH3), increases with decreasing current density, and at j(electrolysis) = 200, 25, 2, and 0.7 mA cm(-2), ?(NH3) = 1%, 7%, 37%, and 71%, respectively. At 200 mA cm(-2), over 90% of applied current drives H2, rather than NH3, formation. Lower temperature supports greater electrolyte hydration. At 105 °C in the hydrated Na(0.5)K(0.5)OH electrolyte, ?(NH3) increases and then is observed to be highly stable at ?(NH3) = 24(+2)%.
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Ammonia synthesis. Ammonia synthesis by N? and steam electrolysis in molten hydroxide suspensions of nanoscale Fe?O?.
Science
PUBLISHED: 08-09-2014
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The Haber-Bosch process to produce ammonia for fertilizer currently relies on carbon-intensive steam reforming of methane as a hydrogen source. We present an electrochemical pathway in which ammonia is produced by electrolysis of air and steam in a molten hydroxide suspension of nano-Fe2O3. At 200°C in an electrolyte with a molar ratio of 0.5 NaOH/0.5 KOH, ammonia is produced at 1.2 volts (V) under 2 milliamperes per centimeter squared (mA cm(-2)) of applied current at coulombic efficiency of 35% (35% of the applied current results in the six-electron conversion of N2 and water to ammonia, and excess H2 is cogenerated with the ammonia). At 250°C and 25 bar of steam pressure, the electrolysis voltage necessary for 2 mA cm(-2) current density decreased to 1.0 V.
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Poly(Ethylene glycol) as a scaffold for high-affinity open-channel blockers of the mouse nicotinic acetylcholine receptor.
PLoS ONE
PUBLISHED: 01-01-2014
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High-affinity blockers for an ion channel often have complex molecular structures that are synthetically challenging and/or laborious. Here we show that high-affinity blockers for the mouse nicotinic acetylcholine receptor (AChR) can be prepared from a structurally simple material, poly(ethylene glycol) (PEG). The PEG-based blockers (PQ1-5), comprised of a flexible octa(ethylene glycol) scaffold and two terminal quaternary ammonium groups, exert low- to sub-micromolar affinities for the open AChR pore (measured via single-channel analysis of AChRs expressed in human embryonic kidney cells). PQ1-5 are comparable in pore-binding affinity to the strongest AChR open-channel blockers previously reported, which have complex molecular structures. These results suggest a general approach for designing potent open-channel blockers from a structurally flexible polymer. This design strategy involves simple synthetic procedures and does not require detailed information about the structure of an ion-channel pore.
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Fabrication of VB2/air cells for electrochemical testing.
J Vis Exp
PUBLISHED: 08-22-2013
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A technique to investigate the properties and performance of new multi-electron metal/air battery systems is proposed and presented. A method for synthesizing nanoscopic VB2 is presented as well as step-by-step procedure for applying a zirconium oxide coating to the VB2 particles for stabilization upon discharge. The process for disassembling existing zinc/air cells is shown, in addition construction of the new working electrode to replace the conventional zinc/air cell anode with a the nanoscopic VB2 anode. Finally, discharge of the completed VB2/air battery is reported. We show that using the zinc/air cell as a test bed is useful to provide a consistent configuration to study the performance of the high-energy high capacity nanoscopic VB2 anode.
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Chemical mechanism of the high solubility pathway for the carbon dioxide free production of iron.
Chem. Commun. (Camb.)
PUBLISHED: 02-08-2011
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We determine the fundamental iron oxide high solubility mechanism that drives a new electrolytic pathway to iron production, and eliminates a major CO(2) emission source, for example it is produced using wind and solar energy, in a molten carbonate electrolyte, at a high rate and a low electrolysis energy.
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High solubility pathway for the carbon dioxide free production of iron.
Chem. Commun. (Camb.)
PUBLISHED: 08-23-2010
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We report a fundamental change in the understanding of iron oxide thermochemistry, opening a facile, new CO(2)-free route to iron production. The resultant process can eliminate a major global source of greenhouse gas emission, producing the staple iron in molten media at high rate and low electrolysis energy.
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Binding Rate Screen - a high-throughput assay in soluble lysate for prioritizing protein expression constructs.
Anal. Biochem.
PUBLISHED: 10-27-2009
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Identification of constructs suitable for the recombinant protein production pipeline is a bottleneck for structural genomics efforts, as most methods require purified proteins and/or are labor-intensive. Here, we present a novel high-throughput approach, Binding Rate Screen, that can alleviate this bottleneck by screening expression constructs in crude soluble lysate. This functional screen utilizes the frequently employed hexahistidine (His(6)) tag as a reporter, and measures its binding rate to an affinity matrix as a metric to reflect aggregation, concentration, and purifiability of the target protein. The constructs with the highest binding rates also exhibit high expression of soluble monomeric protein as judged by analytical size-exclusion chromatography. Constructs expressing variations of the target protein can be prioritized on a time scale of minutes, which is at least 10-100 times faster than any other technologies currently available.
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Functional equivalence of the nicotinic acetylcholine receptor transmitter binding sites in the open state.
Biochim. Biophys. Acta
PUBLISHED: 01-21-2009
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The subunits of the muscle-type nicotinic acetylcholine receptor (AChR) are not uniformly oriented in the resting closed conformation: the two alpha subunits are rotated relative to its non-alpha subunits. In contrast, all the subunits overlay well with one another when agonist is bound to the AChR, suggesting that they are uniformly oriented in the open receptor. This gating-dependent increase in orientational uniformity due to rotation of the alpha subunits might affect the relative affinities of the two transmitter binding sites, making the two affinities dissimilar (functionally non-equivalent) in the initial ligand-bound closed state but similar (functionally equivalent) in the open state. To test this hypothesis, we measured single-channel activity of the alphaG153S gain-of-function mutant receptor evoked by choline, and estimated the resting closed-state and open-state affinities of the two transmitter binding sites. Both model-independent analyses and maximum-likelihood estimation of microscopic rate constants indicate that channel opening makes the binding sites affinities more similar to each other. These results support the hypothesis that open-state affinities to the transmitter binding sites are primarily determined by the alpha subunits.
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STEP wastewater treatment: a solar thermal electrochemical process for pollutant oxidation.
ChemSusChem
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A solar thermal electrochemical production (STEP) pathway was established to utilize solar energy to drive useful chemical processes. In this paper, we use experimental chemistry for efficient STEP wastewater treatment, and suggest a theory based on the decreasing stability of organic pollutants (hydrocarbon oxidation potentials) with increasing temperature. Exemplified by the solar thermal electrochemical oxidation of phenol, the fundamental model and experimental system components of this process outline a general method for the oxidation of environmentally stable organic pollutants into carbon dioxide, which is easily removed. Using thermodynamic calculations we show a sharply decreasing phenol oxidation potential with increasing temperature. The experimental results demonstrate that this increased temperature can be supplied by solar thermal heating. In combination this drives electrochemical phenol removal with enhanced oxidation efficiency through (i) a thermodynamically driven decrease in the energy needed to fuel the process and (ii) improved kinetics to sustain high rates of phenol oxidation at low electrochemical overpotential. The STEP wastewater treatment process is synergistic in that it is performed with higher efficiency than either electrochemical or photovoltaic conversion process acting alone. STEP is a green, efficient, safe, and sustainable process for organic wastewater treatment driven solely by solar energy.
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ATP-sensitive potassium channels exhibit variance in the number of open channels below the limit predicted for identical and independent gating.
PLoS ONE
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In small cells containing small numbers of ion channels, noise due to stochastic channel opening and closing can introduce a substantial level of variability into the cells membrane potential. Negatively cooperative interactions that couple a channels gating conformational change to the conformation of its neighbor(s) provide a potential mechanism for mitigating this variability, but such interactions have not previously been directly observed. Here we show that heterologously expressed ATP-sensitive potassium channels generate noise (i.e., variance in the number of open channels) below the level possible for identical and independent channels. Kinetic analysis with single-molecule resolution supports the interpretation that interchannel negative cooperativity (specifically, the presence of an open channel making a closed channel less likely to open) contributes to the decrease in noise. Functional coupling between channels may be important in modulating stochastic fluctuations in cellular signaling pathways.
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STEP cement: Solar Thermal Electrochemical Production of CaO without CO(2) emission.
Chem. Commun. (Camb.)
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New molten salt chemistry allows solar thermal energy to drive calcium oxide production without any carbon dioxide emission. This is accomplished in a one pot synthesis, and at lower projected cost than the existing cement industry process, which after power production, is the largest contributor to anthropogenic greenhouse gas emissions.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.

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In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.