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Photonic crystals (PCs) are promising tools for label-free sensing in drug discovery screening, diagnostics, and analysis of ligand-receptor interactions. Imaging of PC surface modes has emerged as a novel approach to the detection of multiple binding events at the sensor surface. PC surface modification and decoration with recognition units yield an interface providing the highly sensitive detection of cancer biomarkers, antibodies, and oligonucleotides. The RAD51 protein plays a central role in DNA repair via the homologous recombination pathway. This recombinase is essential for the genome stability and its overexpression is often correlated with aggressive cancer. RAD51 is therefore a potential target in the therapeutic strategy for cancer. Here, we report the designing of a PC-based array sensor for real-time monitoring of oligonucleotide-RAD51 recruitment by means of surface mode imaging and validation of the concept of this approach. Our data demonstrate that the designed biosensor ensures the highly sensitive multiplexed analysis of association-dissociation events and detection of the biomarker of DNA damage using a microfluidic PC array. The obtained results highlight the potential of the developed technique for testing the functionality of candidate drugs, discovering new molecular targets and drug entities. This paves the way to further adaption and bioanalytical use of the biosensor for high-content screening to identify new DNA repair inhibitor drugs targeting the RAD51 nucleoprotein filament or to discover new molecular targets.
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Anticorpos , Neoplasias , Humanos , Diagnóstico por Imagem , Biomarcadores Tumorais , Reparo do DNA , DNA de Cadeia Simples , Oligonucleotídeos , Rad51 RecombinaseRESUMO
8-Oxoguanosine is the most common oxidatively generated base damage and pairs with complementary cytidine within duplex DNA. The 8-oxoguanosine-cytidine lesion, if not recognized and removed, not only leads to G-to-T transversion mutations but renders the base pair being more vulnerable to the ionizing radiation and singlet oxygen (1 O2 ) damage. Herein, reaction dynamics of a prototype Watson-Crick base pair [9MOG â 1MC]â + , consisting of 9-methyl-8-oxoguanine radical cation (9MOGâ + ) and 1-methylcystosine (1MC), was examined using mass spectrometry coupled with electrospray ionization. We first detected base-pair dissociation in collisions with the Xe gas, which provided insight into intra-base pair proton transfer of 9MOGâ + â 1MC â â ${{\stackrel{ {\rightarrow} } { {\leftarrow} } } }$ [9MOG - HN1 ]â â [1MC+HN3' ]+ and subsequent non-statistical base-pair separation. We then measured the reaction of [9MOG â 1MC]â + with 1 O2 , revealing the two most probable pathways, C5-O2 addition and HN7 -abstraction at 9MOG. Reactions were entangled with the two forms of 9MOG radicals and base-pair structures as well as multi-configurations between open-shell radicals and 1 O2 (that has a mixed singlet/triplet character). These were disentangled by utilizing approximately spin-projected density functional theory, coupled-cluster theory and multi-referential electronic structure modeling. The work delineated base-pair structural context effects and determined relative reactivity toward 1 O2 as [9MOG - H]â >9MOGâ + >[9MOG - HN1 ]â â [1MC+HN3' ]+ ≥9MOGâ + â 1MC.
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Citosina , Prótons , Pareamento de Bases , Citosina/química , Cátions , CitidinaRESUMO
Capillary electrophoresis was used to estimate the solvolytic dissociation rate (kd) of metal complexes of deferasirox (DFX, H3L), a drug used to treat iron overload. Inert CoIIIL23- did not dissociate. The estimated kd value for FeIIIL23- was (2.7 ± 0.3) × 10-4 s-1 (298 K, pH 7.4). The kd values of other complexes (AlIIIL23-, NiIIL24-, and MnIIL-) were in the range 10-3-10-4 s-1. In contrast, ZnIIL- and CuIIL- were too labile to allow kd estimation. The fact that the half-life of FeIIIL23- (43.3 min) is shorter than the blood half-life of DFX (8-16 h) implies that the blood concentration of DFX should be high enough to prevent dissociation of FeIIIL23-. The possibility of a safer iron-chelation therapy that avoids excretion of other essential metal ions such as ZnII is discussed, highlighting the importance of selectivity in terms of kinetic stability.
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Sobrecarga de Ferro , Ferro , Humanos , Deferasirox/uso terapêutico , Terapia por Quelação , Quelantes de Ferro , Eletroforese Capilar , BenzoatosRESUMO
Several bifunctional chelators have been synthesized in the last years for the development of new 64Cu-based PET agents for in vivo imaging. When designing a metal-based PET probe, it is important to achieve high stability and kinetic inertness once the radioisotope is coordinated. Different competitive assays are commonly used to evaluate the possible dissociation mechanisms that may induce Cu(II) release in the body. Among them, acid-assisted dissociation tests or transchelation challenges employing EDTA or SOD are frequently used to evaluate both solution thermodynamics and the kinetic behavior of potential metal-based systems. Despite of this, the Cu(II)/Cu(I) bioreduction pathway that could be promoted by the presence of bioreductants still remains little explored. To fill this gap we present here a detailed spectroscopic study of the kinetic behavior of different macrocyclic Cu(II) complexes. The complexes investigated include the cross-bridge cyclam derivative [Cu(CB-TE1A)]+, whose structure was determined using single-crystal X-ray diffraction. The acid-assisted dissociation mechanism was investigated using HClO4 and HCl to analyse the effect of the counterion on the rate constants. The complexes were selected so that the effects of complex charge and coordination polyhedron could be assessed. Cyclic voltammetry experiments were conducted to investigate whether the reduction to Cu(I) falls within the window of common bioreducing agents. The most striking behavior concerns the [Cu(NO2Th)]2+ complex, a 1,4,7-triazacyclononane derivative containing two methylthiazolyl pendant arms. This complex is extremely inert with respect to dissociation following the acid-catalyzed mechanism, but dissociates rather quickly in the presence of a bioreductant like ascorbic acid.
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Complexos de Coordenação , Cobre , Ácido Ascórbico , Quelantes/química , Complexos de Coordenação/química , Cobre/química , Ácido Edético , Ligantes , Tomografia por Emissão de Pósitrons , Superóxido DismutaseRESUMO
A new pyclen-3,9-diacetate derivative ligand (H23,9-OPC2A) was synthesized possessing an etheric O-atom opposite to the pyridine ring, to improve the dissociation kinetics of its Mn(II) complex (pyclen = 3,6,9,15-tetraazabicyclo(9.3.1)pentadeca-1(15),11,13-triene). The new ligand is less basic than the N-containing analogue (H23,9-PC2A) due to the non-protonable O-atom. In spite of its lower basicity, the conditional stability of the [Mn(3,9-OPC2A)] (pMn = -log(Mn(II)), cL = cMn(II) = 0.01 mM. pH = 7.4) remains unaffected (pMn = 8.69), compared to the [Mn(3,9-PC2A)] (pMn = 8.64). The [Mn(3,9-OPC2A)] possesses one water molecule, having a lower exchange rate with bulk solvents (kex298 = 5.3 ± 0.4 × 107 s-1) than [Mn(3,9-PC2A)] (kex298 = 1.26 × 108 s-1). These mild differences are rationalized by density-functional theory (DFT) calculations. The acid assisted dissociation of [Mn(3,9-OPC2A)] is considerably slower (k1 = 2.81 ± 0.07 M-1 s-1) than that of the complexes of diacetates or bisamides of various 12-membered macrocycles and the parent H23,9-PC2A. The [Mn(3,9-OPC2A)] is inert in rat/human serum as confirmed by 52Mn labeling (nM range), as well as by relaxometry (mM range). However, a 600-fold excess of EDTA (pH = 7.4) or a mixture of essential metal ions, propagated some transchelation/transmetalation in 7 days. The H23,9-OPC2A is labeled efficiently with 52Mn at elevated temperatures, yet at 37 °C the parent H23,9-PC2A performs slightly better. Ultimately, the H23,9-OPC2A shows advantageous features for further ligand designs for bifunctional chelators.
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Innovative breakthroughs regarding self-supported open and porous electrodes that can promote gas-liquid transmission and regulate the water dissociation kinetics are critical for sustainable hydrogen economy. Herein, a free-standing porous electrode with Pd-NiS nanoparticles assembled in a multichannel carbonized wood framework (Pd-NiS/CW) was ingeniously constructed. Specifically, carbonized wood (CW) with a mass of open microchannels and high electrical conductivity can significantly facilitate electrolyte permeation ("inhalation"), hydrogen evolution ("exhalation"), and electron transfer. As expected, the fabricated "breathable" wooden electrode exhibits remarkable hydrogen evolution activity in 1.0 M KOH, only requiring a low overpotential of 80 mV to sustain a current density of 10 mA cm-2, and can maintain this current density for 100 h. Further, the spectroscopic characterization and density functional theory (DFT) calculations manifest that the electron interaction between Pd and NiS is beneficial to reduce the water dissociation energy barriers, optimize the adsorption/desorption of H, and ultimately accelerate the catalytic activity. The work reported here will provide a potential approach for the design of electrocatalysts combined with natural multichannel wood to achieve the goal of high electrocatalytic activity and superior durability for hydrogen production.
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The thermodynamic, kinetic, and structural properties of Ln3+ complexes with the bifunctional DO3A-ACE4- ligand and its amide derivative DO3A-BACE4- (modelling the case where DO3A-ACE4- ligand binds to vector molecules) have been studied in order to confirm the usefulness of the corresponding Gd3+ complexes as relaxation labels of targeted MRI contrast agents. The stability constants of the Mg2+ and Ca2+ complexes of DO3A-ACE4- and DO3A-BACE4- complexes are lower than for DOTA4- and DO3A3-, while the Zn2+ and Cu2+ complexes have similar and higher stability than for DOTA4- and DO3A3- complexes. The stability constants of the Ln(DO3A-BACE)- complexes increase from Ce3+ to Gd3+ but remain practically constant for the late Ln3+ ions (represented by Yb3+). The stability constants of the Ln(DO3A-ACE)4- and Ln(DO3A-BACE)4- complexes are several orders of magnitude lower than those of the corresponding DOTA4- and DO3A3- complexes. The formation rate of Eu(DO3A-ACE)- is one order of magnitude slower than for Eu(DOTA)-, due to the presence of the protonated amine group, which destabilizes the protonated intermediate complex. This protonated group causes the Ln(DO3A-ACE)- complexes to dissociate several orders of magnitude faster than Ln(DOTA)- and its absence in the Ln(DO3A-BACE)- complexes results in inertness similar to Ln(DOTA)- (as judged by the rate constants of acid assisted dissociation). The 1H NMR spectra of the diamagnetic Y(DO3A-ACE)- and Y(DO3A-BACE)- reflect the slow dynamics at low temperatures of the intramolecular isomerization process between the SA pair of enantiomers, R-Λ(λλλλ) and S-Δ(δδδδ). The conformation of the Cα-substituted pendant arm is different in the two complexes, where the bulky substituent is further away from the macrocyclic ring in Y(DO3A-BACE)- than the amino group in Y(DO3A-ACE)- to minimize steric hindrance. The temperature dependence of the spectra reflects slower ring motions than pendant arms rearrangements in both complexes. Although losing some thermodynamic stability relative to Gd(DOTA)-, Gd(DO3A-BACE)- is still quite inert, indicating the usefulness of the bifunctional DO3A-ACE4- in the design of GBCAs and Ln3+-based tags for protein structural NMR analysis.
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Complexos de Coordenação/química , Espectroscopia de Ressonância Magnética , Propionatos/química , Ácidos/química , Catálise , Íons , Cinética , Ligantes , Prótons , Soluções , TermodinâmicaRESUMO
We report two macrocyclic ligands based on a 1,7-diaza-12-crown-4 platform functionalized with acetate (tO2DO2A2-) or piperidineacetamide (tO2DO2AMPip) pendant arms and a detailed characterization of the corresponding Mn(II) complexes. The X-ray structure of [Mn(tO2DO2A)(H2O)]·2H2O shows that the metal ion is coordinated by six donor atoms of the macrocyclic ligand and one water molecule, to result in seven-coordination. The Cu(II) analogue presents a distorted octahedral coordination environment. The protonation constants of the ligands and the stability constants of the complexes formed with Mn(II) and other biologically relevant metal ions (Mg(II), Ca(II), Cu(II) and Zn(II)) were determined using potentiometric titrations (I = 0.15 M NaCl, T = 25 °C). The conditional stabilities of Mn(II) complexes at pH 7.4 are comparable to those reported for the cyclen-based tDO2A2- ligand. The dissociation of the Mn(II) chelates were investigated by evaluating the rate constants of metal exchange reactions with Cu(II) under acidic conditions (I = 0.15 M NaCl, T = 25 °C). Dissociation of the [Mn(tO2DO2A)(H2O)] complex occurs through both proton- and metal-assisted pathways, while the [Mn(tO2DO2AMPip)(H2O)] analogue dissociates through spontaneous and proton-assisted mechanisms. The Mn(II) complex of tO2DO2A2- is remarkably inert with respect to its dissociation, while the amide analogue is significantly more labile. The presence of a water molecule coordinated to Mn(II) imparts relatively high relaxivities to the complexes. The parameters determining this key property were investigated using 17O NMR (Nuclear Magnetic Resonance) transverse relaxation rates and 1H nuclear magnetic relaxation dispersion (NMRD) profiles.
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The mechanisms involved in the formation/dissociation of methane hydrate confined at the nanometer scale are unraveled using advanced molecular modeling techniques combined with a mesoscale thermodynamic approach. Using atom-scale simulations probing coexistence upon confinement and free energy calculations, phase stability of confined methane hydrate is shown to be restricted to a narrower temperature and pressure domain than its bulk counterpart. The melting point depression at a given pressure, which is consistent with available experimental data, is shown to be quantitatively described using the Gibbs-Thomson formalism if used with accurate estimates for the pore/liquid and pore/hydrate interfacial tensions. The metastability barrier upon hydrate formation and dissociation is found to decrease upon confinement, therefore providing a molecular-scale picture for the faster kinetics observed in experiments on confined gas hydrates. By considering different formation mechanisms-bulk homogeneous nucleation, external surface nucleation, and confined nucleation within the porosity-we identify a cross-over in the nucleation process; the critical nucleus formed in the pore corresponds either to a hemispherical cap or to a bridge nucleus depending on temperature, contact angle, and pore size. Using the classical nucleation theory, for both mechanisms, the typical induction time is shown to scale with the pore volume to surface ratio and hence the pore size. These findings for the critical nucleus and nucleation rate associated with such complex transitions provide a means to rationalize and predict methane hydrate formation in any porous media from simple thermodynamic data.
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Escherichia coli ClpXP is one of the most thoroughly studied AAA+ proteases, but relatively little is known about the reactions that allow it to bind and then engage specific protein substrates before the adenosine triphosphate (ATP)-fueled mechanical unfolding and translocation steps that lead to processive degradation. Here, we employ a fluorescence-quenching assay to study the binding of ssrA-tagged substrates to ClpXP. Polyphasic stopped-flow association and dissociation kinetics support the existence of at least three distinct substrate-bound complexes. These kinetic data fit well to a model in which ClpXP and substrate form an initial recognition complex followed by an intermediate complex and then, an engaged complex that is competent for substrate unfolding. The initial association and dissociation steps do not require ATP hydrolysis, but subsequent forward and reverse kinetic steps are accelerated by faster ATP hydrolysis. Our results, together with recent cryo-EM structures of ClpXP bound to substrates, support a model in which the ssrA degron initially binds in the top portion of the axial channel of the ClpX hexamer and then is translocated deeper into the channel in steps that eventually pull the native portion of the substrate against the channel opening. Reversible initial substrate binding allows ClpXP to check potential substrates for degrons, potentially increasing specificity. Subsequent substrate engagement steps allow ClpXP to grip a wide variety of sequences to ensure efficient unfolding and translocation of almost any native substrate.
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Endopeptidase Clp/metabolismo , Proteínas de Escherichia coli/metabolismo , ATPases Associadas a Diversas Atividades Celulares/metabolismo , Adenosina Trifosfatases/metabolismo , Trifosfato de Adenosina/metabolismo , Microscopia Crioeletrônica/métodos , Escherichia coli/enzimologia , Escherichia coli/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Hidrólise , Cinética , Dobramento de Proteína , Especificidade por SubstratoRESUMO
Material instability issues, especially moisture degradation in ambient operating environments, limit the practical application of hybrid perovskite in photovoltaic and light-emitting devices. Very recent experiments demonstrate that ligand passivation can effectively improve the surface moisture tolerance of hybrid perovskites. In this work, the interfacial stability of as-synthesized pristine and alkylammonium-passivated methylammonium lead iodide (MAPbI3) with liquid water is systematically investigated using molecular dynamics simulations and reaction kinetics models. Interestingly, the more hydrophilic [PbI2]0 surface is more stable than the less hydrophilic [MAI]0 surface because of the higher polarity of the former surface. Linear alkylammoniums significantly stabilize the [MAI]0 surface with highly reduced (by 1-2 orders of magnitude) dissociation rates of both MA+ and ligands themselves, while branched ligands, surprisingly, lead to higher dissociation rates as the surface coverage increases. Such anomalous behavior is attributed to the aggregation-assisted dissolution of surfactant-like ligands as micelles during the degradation process. Short-chain linear alkylammonium at the full surface coverage is found to be the optimal ligand to stabilize the [MAI]0 surface. This work not only provides fundamental insights into the ionic dissolution pathways and mechanisms of hybrid perovskites in water but also inspires the design of highly stable hybrid perovskites with ligand passivation layers. The computational framework developed here is also transferrable to the investigation of surface passivation chemistry for weak ionic materials in general.
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Molybdenum disulfide (MoS2) has attracted much attention as a promising alternative to Pt-based catalysts for highly efficient hydrogen generation. However, it suffers sluggish kinetics for driving the hydrogen evolution reaction (HER) process because of inert basal planes, especially in alkaline solution. Here, we show a combination of heteroatom doping and phase transformation strategies to engineer the in-plane structure of MoS2, that trigger their catalytic activities. Systematic characterizations are performed with advanced aberration-corrected microscopy and X-ray techniques, indicating that an as-designed MoS2 catalyst has a distorted zigzag-chain superlattice in metallic phase, while its in-plane structure was engineered via the incorporation of cobalt and oxygen species. The optimal Co, O dual-doped metallic phase molybdenum disulfide (1T-MoS2) electrocatalyst shows a significantly enhanced HER activity with a low overpotential of 113 mV at 10 mA cm-2 and corresponding small Tafel slope of 50 mV dec-1, accompanied by the robust stability in alkaline media. The calculated turnover frequency is higher than 6.65 H2 s-1 at an overpotential of 200 mV. More in-depth insights from the first-principle calculations illustrate that the water dissociation as a rate-determining step was largely accelerated by the in-plane Co-O-Mo species and fast electron transfer of the catalyst. Benefiting from ingenious design and fine identifications, this work provides a fundamental understanding of the relationships among heteroatom doping, phase transformation, and performance for MoS2-based catalysts.
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Variable temperature electrospray ionization (ESI) is coupled with mass spectrometry techniques in order to investigate structural transitions of monoclonal antibody immunoglobulin G (IgG) in a 100-mM ammonium acetate (pH 7.0) solution from 26 to 70 °C. At 26 °C, the mass spectrum for intact IgG shows six charge states + 22 to + 26. Upon increasing the solution temperature, the fraction of low-charge states decreases and new, higher-charge state ions are observed. Upon analysis, it appears that heating the solution aids in desolvation of the intact IgG precursor. Above ~ 50 °C, a cleavage event between the light and heavy chains is observed. An analysis of the kinetics for these processes at different temperatures yields transition state thermochemistry of ΔH = 95 ± 10 kJ mol-1, ΔS = 8 ± 1 J mol-1 K-1, and ΔG = 92 ± 11 kJ mol-1. The mechanism for light chain dissociation appears to involve disulfide bond scrambling that ultimately results in a non-native Cys199-Cys217 disulfide bond in the light chain product. Above ~ 70 °C, we are unable to produce a stable ESI signal. The loss of signal is ascribed to aggregation that is primarily associated with the remaining portion of the antibody after having lost the light chain. Graphical Abstract.
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Imunoglobulina G/análise , Imunoglobulina G/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Temperatura Alta , Cinética , Modelos MolecularesRESUMO
A capillary electrophoretic reactor was used to analyze the dissociation kinetics of an enzyme-inhibitor complex in a homogeneous solution without immobilization. The complex consisting of trypsin (Try) and aprotinin (Apr) was used as the model. Capillary electrophoresis provided a reaction field for Try-Apr complex to dissociate through the steady removal of free Try and Apr from the Try-Apr zone. By analyzing the dependence of peak height of Try-Apr on separation time, the dissociation rate kdH was obtained as 2.73â¯×â¯10-4â¯s-1 (298â¯K) at pH 2.46. The dependence of kdH on the proton concentration (pHâ¯=â¯2.09-3.12) revealed a first-order dependence of kdH on [H+]; kdHâ¯=â¯kd + k1[H+], where kd is the spontaneous dissociation rate and was 5.65â¯×â¯10-5â¯s-1, and k1 is the second-order rate constant and was 5.07â¯×â¯10-2â¯M-1â¯s-1. From the kd value, the half-life of the Try-Apr complex at physiological pH was determined as 3.4â¯h. The presence of the proton-assisted dissociation can be explained by the protonation of -COO- of the Asp residue in Try, which breaks the salt bridge with the -NH3+ group of Lys in Apr.
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Aprotinina/química , Inibidores da Tripsina/química , Tripsina/química , Animais , Ligação Competitiva , Bovinos , Eletroforese Capilar , Meia-Vida , Concentração de Íons de Hidrogênio , Cinética , Ligação Proteica , Ressonância de Plasmônio de SuperfícieRESUMO
We report ion mobility spectrometry and mass spectrometry studies of the non-enzymatic step-by-step degradation of substance P (subP), an 11-residue neuropeptide, with the sequence Arg1-Pro2-Lys3-Pro4-Gln5-Gln6-Phe7-Phe8-Gly9-Leu10-Met11-NH2, in ethanol. At elevated solution temperatures (55 to 75 °C), several reactions are observed, including a protonation event, i.e., [subP+2H]2+ + H+ â [subP+3H]3+, that appears to be regulated by a configurational change and two sequential bond cleavages (the Pro2-Lys3 peptide bond is cleaved to form the smaller nonapeptide Lys3-Met11-NH2 [subP(3-11)], and subsequently, subP(3-11) is cleaved at the Pro4-Gln5 peptide bond to yield the heptapeptide Gln5-Met11-NH2 [subP(5-11)]). Each of the product peptides [subP(3-11) and subP(5-11)] is accompanied by a complementary diketopiperazine (DKP): cyclo-Arg1-Pro2 (cRP) for the first cleavage, and cyclo-Lys3-Pro4 (cKP) for the second. Insight about the mechanism of degradation is obtained by comparing kinetics calculations of trial model mechanisms with experimental data. The best model of our experimental data indicates that the initial cleavage of subP is regulated by a conformational change, likely a transâcis isomerization of the Arg1-Pro2 peptide bond. The subP(3-11) product has a long lifetime (t1/2 ~ 30 h at 55 °C) and appears to transition through several structural intermediates prior to dissociation, suggesting that subP(3-11) is initially formed with a Lys3-trans-Pro4 peptide bond configuration and that slow transâcis isomerization regulates the second bond cleavage event as well. From these data and our model mechanisms, we obtain transition state thermochemistry ranging from ΔH = 41 to 85 kJ mol-1 and ΔS = - 43 to - 157 J mol-1 K-1 for each step in the reaction. Graphical Abstract.
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Etanol/metabolismo , Prolina/metabolismo , Substância P/metabolismo , Sequência de Aminoácidos , Temperatura Alta , Hidrólise , Isomerismo , Cinética , Prolina/química , Proteólise , Substância P/química , TermodinâmicaRESUMO
Nucleosomes restrict the occupancy of most transcription factors (TF) by reducing binding and accelerating dissociation, while a small group of TFs have high affinities to nucleosome-embedded sites and facilitate nucleosome displacement. To understand this process mechanistically, we investigated two Saccharomyces cerevisiae TFs, Reb1 and Cbf1. We show that these factors bind to their sites within nucleosomes with similar binding affinities as to naked DNA, trapping a partially unwrapped nucleosome without histone eviction. Both the binding and dissociation rates of Reb1 and Cbf1 are significantly slower at the nucleosomal sites relative to those for naked DNA, demonstrating that the high affinities are achieved by increasing the dwell time on nucleosomes in order to compensate for reduced binding. Reb1 also shows slow migration rate in the yeast nuclei. These properties are similar to those of human pioneer factors (PFs), suggesting that the mechanism of nucleosome targeting is conserved from yeast to humans.
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Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Proteínas de Ligação a DNA/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Fatores de Transcrição/metabolismo , DNA Fúngico/metabolismo , Histonas/metabolismo , Nucleossomos/metabolismo , Ligação ProteicaRESUMO
Although it is commonly believed that the water-dissociation-related Volmer process is the rate-limiting step for alkaline hydrogen evolution reaction (HER) on Pt-based catalysts, the underlying essence, particularly on the atomic scale, still remains unclear. Herein, it is revealed that the sluggish water-dissociation behavior probably stems from unfavorable orbital orientation and the kinetic issue is successfully resolved via N-induced orbital tuning. Impressively, N modified Pt-Ni nanowires deliver an ultralow overpotential of 13 mV at 10 mA cm-2 , which represents a new benchmark for alkaline HER catalysis. Fine-structural characterization and density functional theory analysis illustrate that the introduced nitrogen can uniquely modulate the electron densities around the Ni sites, and further create empty dz 2 orbitals with superior orientation for water adsorption and activation. More importantly, it is demonstrated that N-induced orbital modulation can generally boost the alkaline HER activities of Pt-Co, Pt-Ni, and Pt-Cu, offering a new perspective for the design of HER catalysts and beyond.
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After the serious nuclear accident at the Fukushima Daiichi Nuclear Power Plant caused by the Great East Japan Earthquake in 2011, the development of feasible, safe, and highly sensitive analytical methods (in terms of low levels of radiation exposure and radioactive waste generation) for radioactive samples, especially actinide (An) ions, represents an important challenge. Here we propose a methodology for selecting appropriate emissive probes for An ions with very low consumption and emission of radioactivity by capillary electrophoresis-laser-induced fluorescence detection (CE-LIF), using a small chemical library of probes with eight different chelating moieties. It was found that the emissive probe L1, which possesses the tetradentate chelating moiety 1,10-phenanthroline-2,9-dicarboxylic acid (PDA), was suitable for detecting uranyl ions. The detection limit for the uranyl-L1 complex using CE-LIF combined with dynamic ternary complexation and on-capillary concentration techniques was determined to be 2.9â¯×â¯10-12â¯M (0.7â¯ppt). No interference from the large excess of matrix metal ions was observed. This method was successfully applied to real radioactive liquid samples collected from nuclear facilities, including the Fukushima Daiichi Nuclear Power Plant. This strategy not only permitted the development of a safe and rapid analytical method but also provided insight into the coordination chemistry of An ion complexes. Specifically, the PDA structure provided substantial kinetic inertness to its uranyl complex; the formation of a ternary complex between uranyl-L1 and carbonate was revealed; and unusual interactions were observed between the π-electron systems of uranyl and the phenanthroline ring, which stabilized the uranyl-PDA interaction.
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Herein, the interaction of a potent anticancer drug (Sanguinarine, SG) with dimyristoyl-l-α-phosphatidylglycerol (DMPG) liposome membrane has been investigated at physiological pH. The spectroscopic fluorescence decay results demonstrate a modification of the photophysics of SG within DMPG-encapsulated state leading to preferential stabilization of the iminium ion over the alkanolamine form. This suggests a key role of electrostatic force underlying the interaction. The complex dependence of the thermodynamic parameters on temperature yields a unique finding of a positive heat capacity change (ΔCp) indicating the signature of hydrophobic hydration. The study also demonstrates the application of ß-cyclodextrin (ßCD) as a prospective host system resulting in release of the DMPG-bound drug. A calorimetric exploration of the DMPG-ßCD interaction reveals an intrinsically complex thermodynamics of the process leading to ΔCpâ¯>â¯0 and thus marking the instrumental role of hydrophobic hydration which follows that the DMPG-ßCD interaction is accompanied with burial of polar molecular surfaces. A systematic investigation of the diffusion of the drug within various microheterogeneous environments by Fluorescence Correlation Spectroscopy (FCS) categorically reinforces our arguments.
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Antineoplásicos/química , Benzofenantridinas/química , Interações Hidrofóbicas e Hidrofílicas , Isoquinolinas/química , Fosfatidilgliceróis/química , Água/química , Humanos , Concentração de Íons de Hidrogênio , Lipossomos/química , Estrutura Molecular , Tamanho da Partícula , Espectrometria de Fluorescência , Propriedades de Superfície , TermodinâmicaRESUMO
Drug-binding kinetics could play important roles in determining the efficacy of drugs and has caught the attention of more drug designers. Using the dissociation of 1H-pyrrolo[2,3-b]-pyridines from the focal adhesion kinase as an example, this work finds that steered molecular dynamics simulations could help screen compounds with long-residence times. It also reveals a two-step mechanism of ligand dissociation resembling the release of ADP from protein kinase A reported earlier. A phenyl group attaching to the pyrrole prolongs residence time by creating a large activation barrier for transition from the bound to the intermediate state when it becomes exposed to the solvent. Principal component analysis shows that ligand dissociation does not couple with large-scale collective motions of the protein involving many of its amino acids. Rather, a small subset of amino acids dominates. Some of these amino acids do not contact the ligands directly along the dissociation pathways and could exert long-range allosteric effects. © 2018 Wiley Periodicals, Inc.