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ispartof: JOURNAL OF THE CHINESE CHEMICAL SOCIETY vol:69 issue:1 pages:120-132 status: published

Polypropylene (PP) is being considered as a suitable alternative to the cross‐linked polyethylene (XLPE) insulation as the latter is prone to the space charge accumulation when subjected to the voltage polarity reversals in line‐commutated converter high voltage direct current transmission. This novel contribution correlates the space charge accumulation with the trap distribution inside XLPE and PP under the polarity reversal condition. The pulse electro‐acoustic and surface potential decay methods are used to measure the space charge and trap distributions, respectively. These measurements are carried at different poling durations (1 and 6 h at each polarity) and cycles of voltage polarity reversal (one and three cycles) at the electric field of 60 kV/mm. The XLPE shows more homo and hetero charge accumulation than PP. Dominant shallow traps in the XLPE are responsible for the space charge accumulation, whereas deep traps with higher trap density inhibit space charge accumulation within the PP. Furthermore, the positive and negative threshold fields for the space charge accumulation are measured. It is observed that the space charge can be easily accumulated under the negative electric field. Interestingly, the space charge decays rapidly inside the XLPE with the increased number of polarity reversal cycle. A valid theoretical model is developed to explain the space charge accumulation inside the XLPE and PP. The field‐assisted ionization plays a key role in the space charge accumulation inside the XLPE.

International audience; An original dynamic model for substrate uptake under transient conditions is established and used to simulate a variety of biological responses to external perturbations. The actual uptake and growth rates, treated as cell properties, are part of the model variables as well as the substrate concentration at the cell-liquid interface. Several regulatory loops inspired by the structure of the glycolytic chain are considered to establish a set of ordinary differential equations. The uptake rate evolves so as to reach an equilibrium between the cell demand and the environmental supply. This model does not contain any of the usual algebraic closure laws relating to the instantaneous uptake, growth rates, and the substrate concentration, nor does it enforce the continuity of mass fluxes at the liquid-cell interface. However, these relationships are found in the steady-state solution. Previously unexplained experimental observations are well reproduced by this model. Also, the model structure is suitable for further coupling with flux-based metabolic models and fluid-flow equations.

Understanding the complex interplay among gasotransmitters is of great significance but remains technically challenging. In this study, we present the design and synthesis of a dually responsive BOD-NH-SC reporter for probing the dynamic and alternating existence of NO and H2 S in living cells. This designed reporter can repeatedly cycle S-nitrosation and transnitrosation reactions when successively treated with NO and H2 S, thus affording the interchange of NIR fluorescence at 645 nm (NO) and NIR II fluorescence at 936 nm (H2 S). In light of this unique fluorescence alternation between two colors, we synthesized water-soluble BOD-NH-SC dots to visualize the intracellular dynamics of NO and H2 S. These molecular probes thus provide a toolbox to elucidate the interplaying roles of NO and H2 S in the complex interaction networks of various signal transduction pathways.

The millisecond timescale dynamics of activated Ras transiently sample a low-populated conformational state that has distinct surface property from the major state and represents a promising target for binding of small-molecule compounds. To avoid the complications of hydrolysis, dynamics and other properties of active Ras have so far been routinely investigated by using non-hydrolyzable GTP analogues, which, however, were previously reported to alter both the kinetics and distribution of the conformational exchange. In this study, we quantitatively measured and validated the internal dynamics of Ras complexed with a slowly hydrolyzable GTP analogue, GTPγS, which increases the lifetime of active Ras by 23 times relative to that of native GTP. It was found that GTPγS, in addition to its better mimicking of the exchange kinetics than the commonly used non-hydrolyzable analogues GppNHp and GppCH2 p, can rigorously reproduce the natural dynamics network in active Ras, thus indicating its fitness for use in the development of allosteric inhibitors.

Mitochondrial dynamics (fission and fusion) plays an important role in cell functions. Disruption in mitochondrial dynamics has been associated with diseases such as neurobiological disorders and cardiovascular diseases. Analysis of mitochondrial fission/fusion has been mostly achieved through direct visualization of the fission/fusion events in live-cell imaging of fluorescently labeled mitochondria. In this study, we demonstrated a label-free, non-invasive Electrical Impedance Spectroscopy (EIS) approach to analyze mitochondrial dynamics in a genetically modified human neuroblastoma SH-SY5Y cell line with no huntingtin protein expression. Huntingtin protein has been shown to regulate mitochondria dynamics. We performed EIS studies on normal SH-SY5Y cells and two independent clones of huntingtin-null cells. The impedance data was used to determine the suspension conductivity and further cytoplasmic conductivity and relate to the abnormal mitochondrial dynamics. For instance, the cytoplasm conductivity value was increased by 11% from huntingtin-null cells to normal cells. Results of this study demonstrated that EIS is sensitive to characterize the abnormal mitochondrial dynamics that can be difficult to quantify by the conventional microscopic method.

Drug discovery, in particular optimization of candidates using medicinal chemistry, is generally guided by structural biology. However, for optimizing binding kinetics, relevant for efficacy and off‐target effects, information on protein motion is important. Herein, we demonstrate for the prototypical textbook example of an allegedly “rigid protein” that substantial active‐site dynamics have generally remained unrecognized, despite thousands of medicinal‐chemistry studies on this model over decades. Comparing cryogenic X‐ray structures, solid‐state NMR on micro‐crystalline protein at room temperature, and solution NMR structure and dynamics, supported by MD simulations, we show that under physiologically relevant conditions the pocket is in fact shaped by pronounced open/close conformational‐exchange dynamics. The study, which is of general significance for pharmacological research, evinces a generic pitfall in drug discovery routines., Human carbonic anhydrase II (hCAII) plays an important role as a rigid model protein in medicinal chemistry. Under physiological conditions in solution, NMR spectroscopy combined with molecular dynamics simulations demonstrates that the active site shows pronounced open/close conformational exchange, undetected in crystallography‐based work.

We present an atomistic simulation scheme for the determination of the hydration number (h) of aqueous electrolyte solutions based on the calculation of the water dipole reorientation dynamics. In this methodology, the time evolution of an aqueous electrolyte solution generated from ab initio molecular dynamics simulations is used to compute the reorientation time of different water subpopulations. The value of h is determined by considering whether the reorientation time of the water subpopulations is retarded with respect to bulk‐like behavior. The application of this computational protocol to magnesium chloride (MgCl2) solutions at different concentrations (0.6–2.8 mol kg−1) gives h values in excellent agreement with experimental hydration numbers obtained using GHz‐to‐THz dielectric relaxation spectroscopy. This methodology is attractive because it is based on a well‐defined criterion for the definition of hydration number and provides a link with the molecular‐level processes responsible for affecting bulk solution behavior. Analysis of the ab initio molecular dynamics trajectories using radial distribution functions, hydrogen bonding statistics, vibrational density of states, water‐water hydrogen bonding lifetimes, and water dipole reorientation reveals that MgCl2 has a considerable influence on the hydrogen bond network compared with bulk water. These effects have been assigned to the specific strong Mg‐water interaction rather than the Cl‐water interaction., Ab initio molecular dynamics simulations are used to characterize the structural, vibrational, and dynamical properties of water in aqueous magnesium chloride (MgCl2) solutions, and to develop a methodology to compute the hydration number (h) of aqueous electrolytes from water reorientation dynamics. The application of this protocol gives h values of MgCl2 solutions in excellent agreement with experimental hydration numbers obtained using GHz‐to‐THz dielectric relaxation spectroscopy.

Conformationally flexible protein complexes represent a major challenge for structural and dynamical studies. We present herein a method based on a hybrid NMR/MD approach to characterize the complex formed between the disordered p53TAD1-60 and the metastasis-associated S100A4. Disorder-to-order transitions of both TAD1 and TAD2 subdomains upon interaction is detected. Still, p53TAD1-60 remains highly flexible in the bound form, with residues L26, M40, and W53 being anchored to identical hydrophobic pockets of the S100A4 monomer chains. In the resulting "fuzzy" complex, the clamp-like binding of p53TAD1-60 relies on specific hydrophobic anchors and on the existence of extended flexible segments. Our results demonstrate that structural and dynamical NMR parameters (cumulative Δδ, SSP, temperature coefficients, relaxation time, hetNOE) combined with MD simulations can be used to build a structural model even if, due to high flexibility, the classical solution structure calculation is not possible.

The metallation of nucleic acids is key to wide‐ranging applications, from anticancer medicine to nanomaterials, yet there is a lack of understanding of the molecular‐level effects of metallation. Here, we apply single‐molecule fluorescence methods to study the reaction of an organo‐osmium anticancer complex and DNA. Individual metallated DNA hairpins are characterized using Förster resonance energy transfer (FRET). Although ensemble measurements suggest a simple two‐state system, single‐molecule experiments reveal an underlying heterogeneity in the oligonucleotide dynamics, attributable to different degrees of metallation of the GC‐rich hairpin stem. Metallated hairpins display fast two‐state transitions with a two‐fold increase in the opening rate to ~2 s‐1, relative to the unmodified hairpin, and relatively static conformations with long‐lived open (and closed) states of 5 s to ≥ 50 s. These studies show that a single‐molecule approach can provides new insight into metallation‐induced changes in DNA structure and dynamics.

Nifedipine is calcium channels and pumps blocker widely used in medicine. However, mechanisms of nifedipine action in blood are not clear. In particular, the influence of nifedipine on erythrocytes is far from completely understood. In this work, applying scanning flow cytometry, we observed experimentally for the first time the dynamics behind a significant increase of HCO3 - /Cl- transmembrane exchange rate of CDB3 (main anion exchanger, AE1, Band 3, SLC4A1) of human erythrocytes in the presence of nifedipine in blood. It was found that the rate of CDB3 activation is not limited by the rate of nifedipine binding and/or Ca2+ transport. In order to explain the experimental data, we suggested a kinetic model assuming that the rate of CDB3 activation is limited by the dynamics of the balance between two intracellular processes (1) the activation of CDB3 limited by its interaction with intracellular Ca2+ , and (2) the spontaneous deactivation of CDB3. Thus the use of scanning flow cytometry allowed to clarify quantitatively the molecular kinetic mechanism of nifedipine action on human erythrocytes. In particular, the efficiency (~30) and rates of activation (~0.3 min-1 ) and deactivation (~10-3 min-1 ) of CDB3 in human erythrocytes was evaluated for two donors. © 2019 International Society for Advancement of Cytometry.

Glutamine-binding protein (GlnBP) from Escherichia coli is a prototypical periplasmic binding protein that has been crystallized in an apo, “open” conformation, with its two domains far apart, and a holo, “closed” form, with proximal domains that engulf the cognate L-Gln ligand. A fundamental question about such large-scale conformational transitions—whether the closed state exists in the absence of ligand—is a matter of controversy in the case of GlnBP. Previously, NMR observations have indicated no evidence of the closed form, whereas experimentally validated computations have suggested a remarkable ~40% population. Here, a paramagnetic NMR strategy designed to specifically detect the putative apo-closed species shows that a major population of the latter is highly improbable. Further, NMR residual dipolar couplings collected under three anisotropic conditions fail to reveal differential domain alignment (which would otherwise signal interdomain dynamics) and establish that the average solution conformation is satisfied by the apo-open crystal structure. Our results indicate that the computational prediction of large-scale interdomain motions is far from trivial and may lead to grossly erroneous conclusions without proper experimental validation.