Binding ability of a HHP-tagged protein towards Ni2+ studied by paramagnetic NMR relaxation: The possibility of obtaining long-range structure information

The binding ability of a protein with a metal binding tag towards Ni(2+) was investigated by longitudinal paramagnetic NMR relaxation, and the possibility of obtaining long-range structure information from the paramagnetic relaxation was explored. A protein with a well-defined solution structure (Escherichia coli thioredoxin) was used as the model system, and the peptide His-His-Pro (HHP) fused to the N-terminus of the protein was used as the metal binding tag. It was found that the tag forms a stable dimer complex with the paramagnetic Ni(2+) ion, where each metal ion binds two HHP-tagged protein molecules. However, it was also found that additional sites in the protein compete with the HHP-tag for the binding of the metal ion. These binding sites were identified as the side chain carboxylate groups of the aspartic and glutamic acid residues. Yet, the carboxylate groups bind the Ni(2+) ions considerably weaker than the HHP-tag, and only protons spatially close to the carboxylate sites are affected by the Ni(2+) ions bound to these groups. As for the protons that are unaffected by the carboxylate-bound Ni(2+) ions, it was found that the long-range distances derived from the paramagnetic relaxation enhancements are in good agreement with the solution structure of thioredoxin. Specifically, the obtained long-range paramagnetic distance constraints revealed that the dimer complex is asymmetric with different orientations of the two protein molecules relative to the Ni(2+) ion.     

Phytoextraction of Soil Cobalt Using Hyperaccumulator Plants

A greenhouse study was conducted on phytoextraction of cobalt by nickel hyperaccumulators Alyssum murale and Alyssum corsicum and by two varieties of cobalt accumulator Nyssa sylvatica compared with the nonmetal accumulator crop plant Brassica juncea. The plants were grown on Sassafras sandy loam soil (<2 mg Co and 5 mg Ni/kg dry soil), amended with 1 mmol Co/kg dry soil (58.9 mg/kg), and two Ni smelter-contaminated soils, Quarry muck with 24 mg Co and 1720 mg Ni/kg dry soil and Welland loam with 37 mg Co and 2570 mg Ni/kg dry soil. All soils were adjusted to pH 6.5 to prevent Ni phytotoxicity. Of the five plant entries tested in the study, the two Alyssum species demonstrated the most promising Co phytoextraction results. In Co-amended Sassafras soil, the maximum concentration accumulated by Alyssum murale was 1320 mg Co/kg dry weight, which was almost 60 times higher than accumulation by crop plant Brassica juncea. At a single harvest after 60 days of growth, A. murale was able to extract more than 3% of Co from Co-amended soil. As expected, both Alyssum species accumulated up to 1% Ni on dry weight basis when grown on Ni-contaminated soils.

Nyssa sylvatica showed considerable Co accumulation; foliar Co concentration in the second harvest was as high as 800 mg/kg dry weight. The first few leaves that emerged were chlorotic, both in the Co-amended soil and Ni-contaminated soils, but with growth the signs of toxicity disappeared. In the Co amended soil, Co concentration in Nyssa sylvatica leaves was 30% of that found in shoots of Alyssum species, but an order of magnitude higher than that of Brassica juncea. The leaves accumulated a higher concentration compared with the stems.

Both Alyssum species and Nyssa sylvatica offer promise for phytoextraction of Co and ⁶⁰Co from contaminated or mineralized soils.     

Effects of nickel hyperaccumulation on physiological characteristics of Alyssum murale grown on metal contaminated waste amended soil

A pot experiment was conducted to investigate the effect of nickel concentration on physiological characteristics of Alyssum murale when grown in a soil mixed with sewage sludge (at the rate of 2.8%). Two types of sludge were used: agricultural sewage sludge (S1) and industrial sewage sludge with an increasing nickel concentration (S2, S3, and S4). Results showed that Ni in shoots was higher than Ni in roots. A. murale is able to concentrate up to 12730 mg/kg Ni in leaves. The highest dry matter yield was observed with plants grown with agricultural sewage sludge. An addition of S2 and S3 increased shoot biomass. However, application of S4 reduced 40% shoot dry weight as compared to the control Ni treatment did not affect all chlorophyll fluorescence parameters. The F(v)/F(m) ratio was stable between Ni treatments. Photosynthesis rate (A) increased with agricultural sewage sludge, but remained stable with variable Ni rates from the industrial sludge. The chlorophyll content increased with S1, S2 and S3 but it remains constant with S4 when compared to the control Therefore, high nickel concentration did not affect the function of the photosynthetic machine of A. murale.     

Selective blockage of Serratia marcescens ShlA by nickel inhibits the pore‐forming toxin‐mediated phenotypes in eukaryotic cells

Serratia marcescens is an opportunistic pathogen with increasing incidence in clinical settings. This is mainly attributed to the timely expression of a wide diversity of virulence factors and intrinsic and acquired resistance to antibiotics, including β‐lactams, aminoglycosides, quinolones, and polypeptides. For these reasons, S. marcescens has been recently categorised by the World Health Organization as one priority to strengthen efforts directed to develop new antibacterial agents. Therefore, it becomes critical to understand the underlying mechanisms that allow Serratia to succeed within the host. S. marcescens ShlA pore‐forming toxin mediates phenotypes that alter homeostatic and signal transduction pathways of host cells. It has been previously demonstrated that ShlA provokes cytotoxicity, haemolysis and autophagy and also directs Serratia egress and dissemination from invaded nonphagocytic cells. However, molecular details of ShlA mechanism of action are still not fully elucidated. In this work, we demonstrate that Ni²⁺ selectively and reversibly blocks ShlA action, turning wild‐type S. marcescens into a shlA mutant strain phenocopy. Combined use of Ni²⁺ and calcium chelators allow to discern ShlA‐triggered phenotypes that require intracellular calcium mobilisation and reveal ShlA function as a calcium channel, providing new insights into ShlA mode of action on target cells.     

A Pair of NiCo2O4 and V2O5 Nanowires Directly Grown on Carbon Fabric for Highly Bendable Lithium‐Ion Batteries

Mechanically bendable and flexible functionalities are urgently required for next‐generation battery systems that will be included in soft and wearable electronics, active sportswear, and origami‐based deployable space structures. However, it is very difficult to synthesize anode and cathode electrodes that have high energy density and structural reliability under large bending deformation. Here, vanadium oxide (V₂O₅) and nickel cobalt oxide (NiCo₂O₄) nanowire‐carbon fabric electrodes for highly flexible and bendable lithium ion batteries are reported. The vanadium oxide and nickel cobalt oxide nanowires were directly grown on plasma‐treated carbon fabric and were used as cathode and anode electrodes in a full cell lithium ion battery. Most importantly, a pre‐lithiation process was added to the nickel cobalt oxide nanowire anode to facilitate the construction of a full cell using symmetrically‐architectured nanowire‐carbon fabric electrodes. The highly bendable full cell based on poly(ethylene oxide) polymer electrolyte and room temperature ionic liquid shows high energy density of 364.2 Wh kg^?1 at power density of 240 W kg^?1, without significant performance degradation even under large bending deformations. These results show that vanadium oxide and lithiated nickel cobalt oxide nanowire‐carbon fabrics are a good combination for binder‐free electrodes in highly flexible lithium‐ion batteries.     

Copper–Nickel Nitride Nanosheets as Efficient Bifunctional Catalysts for Hydrazine‐Assisted Electrolytic Hydrogen Production

Electrocatalytic water splitting is one of the sustainable and promising strategies to generate hydrogen fuel but still remains a great challenge because of the sluggish anodic oxygen evolution reaction (OER). A very effective approach to dramatically decrease the input cell voltage of water electrolysis is to replace the anodic OER with hydrazine oxidation reaction (HzOR) due to its lower thermodynamic oxidation potential. Therefore, developing the low‐cost and efficient HzOR catalysts, coupled with the cathodic hydrogen evolution reaction (HER), is tremendously important for energy‐saving electrolytic hydrogen production. Herein, a new‐type of copper–nickel nitride (Cu₁Ni₂‐N) with rich Cu₄N/Ni₃N interface is rationally constructed on carbon fiber cloth. The 3D electrode exhibits extraordinary HER performance with an overpotential of 71.4 mV at 10 mA cm^?2 in 1.0 m KOH, simultaneously delivering an ultralow potential of 0.5 mV at 10 mA cm^?2 for HzOR in a 1.0 m KOH/0.5 m hydrazine electrolyte. Moreover, the electrolytic cell utilizing the synthesized Cu₁Ni₂‐N electrode as both the cathode and anode display a cell voltage of 0.24 V at 10 mA cm^?2 with an excellent stability over 75 h. The present work develops the promising copper–nickel‐based nitride as a bifunctional electrocatalyst through hydrazine‐assistance for energy‐saving electrolytic hydrogen production.     

Investigation of Vetiver Grass Capability in Phytoremediation of Contaminated Soils with Heavy Metals (Pb,Cd, Mn,and Ni)

ABSTRACT

Soil contamination with heavy metals has become a worldwide concern. A sustainable technology to mitigate heavy metal contamination is extremely important. Phytoremediation is a cost-effective method, environmentally friendly, and esthetically pleasing. The aim of this study was to investigate the potential of Vetiver phytoremediation of soils contaminated with heavy metals. This research was conducted as a factorial design with four different heavy metals (lead, cadmium, manganese, and nickel) with three varying levels and also three replications for each treatment. Statistical analysis of data was performed using SPSS19 software and analysis of variance, Duncan and Pearson correlation tests. The results showed that, the highest uptake rate was related to lead metal with 282.45 mg/kg of dry soil and 83.4% uptake percentage. Then, the mean and percentage of adsorption for cadmium, nickel and manganese were 248.3 mg/kg (53.2%), 69.4 mg/kg (65.5%), and 63.29 mg/kg (61%), respectively. Lead was found to be the main component of uptake by Vetiver plant. It was found that the roots of the plant have absorbed more heavy metals than the shoots. And at the roots in total 1089.05 and on average 363.01 mg/kg and at the shoots 901.19 and on average 300.39 mg/kg, the metals used were adsorbed on three levels and four treatments. The results of analysis of variance, Duncan test and Pearson correlation showed that the effect of applied treatments on lead uptake in roots and shoots increased significantly (P ≤ 0.05) with increasing levels of treatments. The biological concentration factor was more than one, and the transfer factor was close to one. Therefore, it can be used as a phytostablization plant. The results showed that Vetiver can be considered as a refining plant due to its vegetative characteristics, cost-effectiveness and high adaptation to environmental conditions.     

Reduced cellular Ca availability enhances TDP-43 cleavage by apoptotic caspases

Accumulation of transactive response DNA binding protein (TDP-43) fragments in motor neurons is a post mortem hallmark of different neurodegenerative diseases. TDP-43 fragments are the products of the apoptotic caspases-3 and -7. Either excessive or insufficient cellular Ca²⁺ availability is associated with activation of apoptotic caspases. However, as far as we know, it is not described whether activation of caspases, due to restricted intracellular Ca²⁺, affects TDP-43 cleavage. Here we show that in various cell lineages with restricted Ca²⁺ availability, TDP-43 is initially cleaved by caspases-3 and -7 and then, also by caspases-6 and -8 once activated by caspase-3. Furthermore, we disclose the existence of a TDP-43 caspase-mediated fragment of 15 kDa, in addition to the well-known fragments of 35 and 25 kDa. Interestingly, with respect to the other two fragments this novel fragment is the major product of caspase activity on murine TDP-43 whereas in human cell lines the opposite occurs. This outcome should be considered when murine models are used to investigate TDP-43 proteinopathies.     

Residue specific partitioning of KL4 into phospholipid bilayers

KL₄, which has demonstrated success in the treatment of respiratory distress, is a synthetic helical, amphipathic peptide mimetic of lung surfactant protein B. The unusual periodicity of charged residues within KL₄ and its relatively high hydrophobicity distinguish it from canonical amphipathic helical peptides. Here we utilized site specific spin labeling of both lipids and the peptide coupled with EPR spectroscopy to discern the effects of KL₄ on lipid dynamics, the residue specific dynamics of hydrophobic regions within KL₄, and the partitioning depths of specific KL₄ residues into the DPPC/POPG and POPC/POPG lipid bilayers under physiologically relevant conditions. KL₄ induces alterations in acyl chain dynamics in a lipid-dependent manner, with the peptide partitioning more deeply into DPPC-rich bilayers. Combined with an earlier NMR study of changes in lipid dynamics on addition of KL₄ (V.C. Antharam et al., 2009), we are able to distinguish how KL₄ affects both collective bilayer motions and intramolecular acyl chain dynamics in a lipid-dependent manner. EPR power saturation results for spin labeled lipids demonstrate that KL₄ also alters the accessibility profiles of paramagnetic colliders in a lipid-dependent manner. Measurements of dynamics and depth parameters for individual spin-labeled residues within KL₄ are consistent with a model where the peptide partitions deeply into the lipid bilayers but lies parallel to the bilayer interface in both lipid environments; the depth of partitioning is dependent on the degree of lipid acyl chain saturation within the bilayer.