G-Quadruplexes as Sensors of Intracellular Na+/K+ Ratio: Potential Role in Regulation of Transcription and Translation.

Data on the structure of G-quadruplexes, noncanonical nucleic acid forms, supporting an idea of their potential participation in regulation of gene expression in response to the change in intracellular Na+i/K+i ratio are considered in the review. Structural variety of G-quadruplexes, role of monovalent cations in formation of this structure, and thermodynamic stability of G-quadruplexes are described. Data on the methods of their identification in the cells and biological functions of these structures are presented. Analysis of information about specific interactions of G-quadruplexes with some proteins was conducted, and their potential participation in the development of some pathological conditions, in particular, cancer and neurodegenerative diseases, is considered. Special attention is given to the plausible role of G-quadruplexes as sensors of intracellular Na+i/K+i ratio, because alteration of this parameter affects folding of G-quadruplexes changing their stability and, thereby, organization of the regulatory elements of nucleic acids. The data presented in the conclusion section demonstrate significant change in the expression of some early response genes under certain physiological conditions of cells and tissues depending on the intracellular Na+i/K+i ratio.

Regulation of Mitochondrial Permeability Transition Pore Opening by Monovalent Cations in Liver Mitochondria.

The opening of the permeability transition pore (PTP) in mitochondria is a key event in the initiation of cell death in various pathologic states, including ischemia/reperfusion. The activation of K+ transport into mitochondria protects cells from ischemia/reperfusion. However, the role of K+ transport in PTP regulation is unclear. Here, we studied the role of K+ and other monovalent cations in the regulation of the PTP opening in an in vitro model. The registration of the PTP opening, membrane potential, Ca2+-retention capacity, matrix pH, and K+ transport was performed using standard spectral and electrode techniques. We found that the presence of all cations tested in the medium (K+, Na+, choline+, and Li+) strongly stimulated the PTP opening compared with sucrose. Several possible reasons for this were examined: the effect of ionic strength, the influx of cations through selective and non-selective channels and exchangers, the suppression of Ca2+/H+ exchange, and the influx of anions. The data obtained indicate that the mechanism of PTP stimulation by cations includes the suppression of K+/H+ exchange and acidification of the matrix, which facilitates the influx of phosphate. Thus, the K+/H+ exchanger and the phosphate carrier together with selective K+ channels compose a PTP regulatory triad, which might operate in vivo.

Markedly Different Effects of Monovalent Cations on the Efficiency of Gene Expression.

The effect of monovalent cations on a cell-free transcription-translation (TX-TL) system is examined using a luciferase assay. It is found that the potency for all ions analyzed here is in the order Rb+  > K+  > Cs+  > Na+  ≈ Li+  > (CH3 )4 N+ , where Rb+ is most efficient at promoting TX-TL and the ions of Li+ , Na+ , and (CH3 )4 N+ exhibit an inhibitory effect. Similar promotion/inhibition effects are observed for cell-free TL alone with an mRNA template.

The influence of pH and monovalent/divalent cations on the structural and physicochemical properties of myofibrillar protein from silver carp.

The present study aimed to investigate effects of pH and monovalent (Na+ and K+)/divalent (Ca2+ and Mg2+) cations on the structural and physicochemical properties of myofibrillar protein (MP) from silver carp. MP treated with divalent cation had lesser change for the structure than that treated with monovalent cation. Ca2+-ATPase activity of MP treated with monovalent cation was increased firstly and then decreased, while that treated with divalent cation was decreased with increasing ionic strength. Surface hydrophobicity and Z-average of MP treated with divalent cations was lower than that with monovalent cations, while they decreased and then increased with the pH shifting from 3.0 to 9.0. Zeta potential of MP was increased and then decreased with increasing the pH but decreased and then increased with increasing ionic strength. In general, the pH and monovalent/divalent cations could cause various hydrophobic and electrostatic interactions, resulting in changes of the physicochemical properties of MP.

Response of anaerobic digestion of waste activated sludge to types of alkalis: Contribution identification of metal ions.

Alkaline pretreatment is one promising strategy for promoting anaerobic digestion of waste activated sludge (WAS). This study selected three types of alkalis with monovalent (NaOH and KOH), divalent (Ca(OH)2 and Mg(OH)2), and trivalent (Fe(OH)3 and Al(OH)3) cations to reveal the roles of metal ions on short chain fatty acids (SCFAs) production. The enhanced production potentials of SCFAs were reduced by order of alkalis with monovalent, divalent, and trivalent cations. Na+, K+, Ca2+, and Mg2+ did no contributions on SCFAs production, while Fe3+ and Al3+ performed better than control, especially the latter. The mechanism analysis proved that Na+, K+, Ca2+, and Mg2+ did no significant effects on solubilization, hydrolysis, acidification and methanogenesis stages, while the first three stages were improved by Fe3+ and Al3+ and the methanogenesis stage was inhibited. The findings may provide some new insights when using alkalis or residual metal ions to improve anaerobic digestion of WAS.

Activation and thermal stabilization of a recombinant γ-glutamyltranspeptidase from Bacillus licheniformis ATCC 27811 by monovalent cations.

The regulation of enzyme activity through complexation with certain metal ions plays an important role in many biological processes. In addition to divalent metals, monovalent cations (MVCs) frequently function as promoters for efficient biocatalysis. Here, we examined the effect of MVCs on the enzymatic catalysis of a recombinant γ-glutamyltranspeptidase (BlrGGT) from Bacillus licheniformis ATCC 27,811 and the application of a metal-activated enzyme to L-theanine synthesis. The transpeptidase activity of BlrGGT was enhanced by Cs+ and Na+ over a broad range of concentrations with a maximum of 200 mM. The activation was essentially independent of the ionic radius, but K+ contributed the least to enhancing the catalytic efficiency. The secondary structure of BlrGGT remained mostly unchanged in the presence of different concentrations of MVCs, but there was a significant change in its tertiary structure under the same conditions. Compared with the control, the half-life (t1/2) of the Cs+-enriched enzyme at 60 and 65 °C was shown to increase from 16.3 and 4.0 min to 74.5 and 14.3 min, respectively. The simultaneous addition of Cs+ and Mg2+ ions exerted a synergistic effect on the activation of BlrGGT. This was adequately reflected by an improvement in the conversion of substrates to L-theanine by 3.3-15.1% upon the addition of 200 mM MgCl2 into a reaction mixture comprising the freshly desalted enzyme (25 µg/mL), 250 mM L-glutamine, 600 mM ethylamine, 200 mM each of the MVCs, and 50 mM borate buffer (pH 10.5). Taken together, our results provide interesting insights into the complexation of MVCs with BlrGGT and can therefore be potentially useful to the biocatalytic production of naturally occurring γ-glutamyl compounds. KEY POINTS: • The transpeptidase activity of B. licheniformis Î³-glutamyltranspeptidase can be activated by monovalent cations. • The thermal stability of the enzyme was profoundly increased in the presence of 200 mM Cs+. • The simultaneous addition of Cs+and Mg2+ions to the reaction mixture improves L-theanine production.

Discrepant effects of monovalent cations on membrane fouling induced by colloidal polymer: Evaluation and mechanism investigation.

Understanding how ionic conditions affect membrane fouling induced by anionic polyacrylamide (APAM) is important for achieving long-term and stable operation of a polymer flooding produced wastewater (PFPW) membrane separation process. However, there is lack of studies on the effects of monovalent cations (Na+ and K+) on APAM-based membrane fouling. In this work, the effects of Na+ and K+ on filtration efficiency, flux decline behavior, fouling resistance, and cleaning efficiency were studied through a series of microfiltration tests. Moreover, the influencing mechanism of membrane fouling was further comprehensively revealed from the aspects of the extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) theory, the hydration force, and the microstructure characterizations. The XDLVO theory agreed well with membrane fouling behavior at various ionic strengths. The increase in ionic strength (0-10,000 mg/L) of Na+ and K+ exacerbated the reduction of relative flux (J/J0) and the accumulation of fouling resistance, as well as made the porous APAM-induced fouling layer denser and more compact, boosting removal efficiency. Furthermore, K+ had a stronger aggravating effect on membrane fouling than Na+. Specifically, the final value of J/J0 for APAM+K+ (0.08) was lower than that for APAM + Na+ (0.12), and the fouling resistance for APAM+K+ (12.25 × 1011 m-1) was higher than that for APAM + Na+ (12.01 × 1011 m-1) at an ionic strength of 10,000 mg/L, which was owing to the larger hydration force caused by Na+ with a smaller ionic radius. This research offers practical guidance for the PFPW membrane filtering process.

BADH-NAD+-K+ Complex Interaction Studies Reveal a New Possible Mechanism between Potassium and Glutamic 254 at the Coenzyme Binding Site.

Betaine aldehyde dehydrogenase (BADH EC 1.2.1.8) catalyzes the irreversible oxidation of betaine aldehyde to glycine betaine using NAD+ as a coenzyme. Incubation of porcine kidney BADH (pkBADH) with NAD+ decreases the catalytic cysteine (C288) reactivity. Potassium ion increases the pkBADH affinity by the coenzyme. This work aimed to analyze pkBADH and NAD+ interaction in the presence and absence of K+ using 1H NMR to identify the amino acids that interact with NAD+ and/or K+ to understand the regulation process of pkBADH-NAD+ complex formation mediated by the K+ ion and their impact on the substrate binding and catalysis. Nuclear magnetic resonance spectra of pkBADH were obtained in the presence and absence of NAD+ and K+. The results show a chemical shift of the signals corresponding to the catalytic glutamic that participates in the transfer of H+ in the reaction of the pkBADH-NAD+-K+ complex formation. Furthermore, there is a widening of the signal that belongs to the catalytic cysteine indicating higher rigidity or less grade of rotation of the structure, which is consistent with the possible conformations of C288 in the catalytic process; in addition, there is evidence of changes in the chemical environment that surrounds NAD+.

Using capillary electrophoresis to characterize the hydrodynamic and electrostatic properties of DNA in solutions containing various monovalent cations.

This review describes the results obtained by using free-solution capillary electrophoresis to probe the electrostatic and hydrodynamic properties of DNA in solutions containing various monovalent cations. In brief, we found that the mobilities of double-stranded DNAs (dsDNAs) increase with increasing molecular weight before leveling off and becoming constant at molecular weights ≥400 bp. The mobilities of single-stranded DNAs (ssDNAs) go through a maximum at ∼10-20 nucleotides before decreasing and becoming constant for oligomers larger than ∼30-50 bases. The mobilities of both ss- and dsDNAs increase linearly with the logarithm of increasing charge per unit length and decrease linearly with the logarithm of increasing ionic strength. Surprisingly, ss- and dsDNA mobilities level off and become nearly constant at ionic strengths ≥0.6 M. The thermal stabilities of dsDNAs decrease linearly with increasing solution viscosity. The diffusion coefficients of dsDNA are modulated by the diffusion coefficients of their counterions because of electrostatic DNA-cation coupling interactions. Finally, the anomalously slow mobilities observed for A-tract-containing DNAs can be attributed both to differences in shape and to the preferential localization of small cations in the A-tract minor groove. Since many of these results are mirrored in other polyion-counterion systems, free-solution electrophoresis can be viewed as a reporter of the electrostatics and hydrodynamics of highly charged polyions. New results describing the mobilities of dsDNA analogues of a microRNA-messenger RNA complex are also presented.

Efficient Low-Grade Heat Harvesting Enabled by Tuning the Hydration Entropy in an Electrochemical System.

Harvesting of low-grade heat (<100 °C) is promising, but its application is hampered by a lack of efficient and low-cost systems. The thermally regenerative electrochemical cycle (TREC) is a potential alternative system with high energy-conversion efficiency. Here, the temperature coefficient (α), which is a key factor in a TREC, is studied by tuning the hydration entropy of the electrochemical reaction. The change of α in copper hexacyanoferrate (CuHCFe) with intercalation of different monovalent cations (Na+ , K+ , Rb+ , and Cs+ ) and a larger α value of -1.004 mV K-1 being found in the Rb+ system are observed. With a view to practical application, a full cell is constructed for low-grade heat harvesting. The resultant ηe is 4.34% when TREC operates between 10 and 50 °C, which further reaches 6.21% when 50% heat recuperation is considered. This efficiency equals to 50% of the Carnot efficiency, which is thought to be the highest ηe reported for low-grade heat harvesting systems. This study provides a fundamental understanding of the mechanisms governing the TREC, and the demonstrated efficient system paves the way for low-grade heat harvesting.

Effects of cations on biofilm formation and characteristics in integrated fixed film activated sludge process at different carbon and nitrogen loadings.

Both divalent cations including calcium and magnesium play important roles for microbial aggregates in binding to negatively charged functional groups on bacterial surfaces, in extracellular polymeric substances (EPS), and on inorganic materials in flocs and biofilms. Monovalent cations such as sodium and potassium deteriorate the floc structure and physical properties. The Integrated Fixed Activated Sludge (IFAS) process employs fixed film media in the aerobic zone; therefore, both monovalent and divalent cations are involved in the process performances. In this study, the effects of cations indicated as the monovalent to divalent cations (M/D) ratio on the biofilm formation and characteristics, and on the IFAS performances for carbon and ammonium removals were evaluated. The experiments were conducted in three IFAS systems feeding with the same wastewater but different M/D ratios and two carbon and nitrogen loadings. The findings revealed that high monovalent with low divalent cations at the M/D ratios higher than 2.0 produced excessive polysaccharides in EPS resulting in high viscosity of activated sludge flocs causing viscous bulking with high SVI values, decreasing the biofilm formation, and increasing the biofilm sloughing. Increasing of both monovalent and carbon loading increased more polysaccharides in the EPS leading to the failures of IFAS system. Nitrification failed at higher M/D ratios because of less nitrifiers in flocs and biofilm. The M/D ratio less than 2.0 is suggested to minimize the excessive EPS production in the IFAS system, especially at high organic loading.

Impacts of mono/divalent cations on the lamellar structure of cross-linked GO layers and membrane filtration performance for different DOM fractions.

This study evaluated the effects of co-existing cations (Na+ or Ca2+) on the lamellar structure of cross-linked graphene oxide (GO) layers and GO modified membrane performance in terms of their fouling behaviours and retention for single-model organic matter, namely, bull serum albumin (BSA), sodium alginate (SA), humic acid (HA) and tannic acid (TA). In the absence of co-existing cations, the GO layers mitigated membrane fouling for large molecules (SA, BSA, and HA) but led to severer pore blocking for small molecules (TA) compared with pristine membrane. Na+ and Ca2+ altered the performance impacts of the GO modified membrane due to different interactions with the cross-linked GO layers. Low concentrations of Na+ (<0.4 mM) enlarged interlayer spacing of the GO layers and caused a decrease in flux after physical cleaning, but the GO layers maintained the uniform lamellar structure. High concentrations of Na+ (>0.4 mM) promoted the aggregation of cross-linked GO layers through charge shielding and reduced the uniformity of lamellar structure, which weakened the antifouling performance for large molecules and promoted the passage of small molecules through the membrane. However, Ca2+ complexed with GO sheets and reinforced the uniform lamellar structure of the GO layers, leading to a better antifouling performance for the filtration of large molecules than the pristine membrane but aggravated TA fouling.

High Selectivities among Monovalent Cations in Dialysis through Cation-Exchange Membranes Coated with Polyelectrolyte Multilayers.

Cation-exchange membranes allow preferential passage of cations over anions, but they show minimal selectivity among cations, which limits their use in ion separations. Recent studies show that modification of cation-exchange membranes with polyelectrolyte multilayers leads to exceptional monovalent/divalent cation electrodialysis selectivities, but no studies report high selectivity among monovalent ions. This work demonstrates that adsorption of protonated poly(allylamine) (PAH)/poly(4-styrenesulfonate) (PSS) multilayers on Nafion membranes leads to high K+/Li+ selectivities in Donnan dialysis, where K+ and Li+ ions in a source phase pass through the membrane and exchange with Na+ ions in a receiving phase. Addition of 0.01 M HNO3 to a source phase containing 0.01 M KNO3 and 0.01 M LiNO3 increases the K+/Li+ selectivity from 8 to ∼60 through (PAH/PSS)5PAH-coated Nafion membranes, primarily because of a ≥fivefold increase in K+ flux. These selectivities are much larger than the ratio of 1.9 for the aqueous diffusion coefficients of K+ and Li+, and uncoated Nafion membranes give a K+/Li+ selectivity <3. Bi-ionic transmembrane potential measurements at neutral pH confirm that the membrane is more permeable to K+ than Li+, but this selectivity is less than in Donnan dialysis with acidic solutions. In situ ellipsometry data indicate that PAH/PSS multilayers (assembled at pH 2.3, 7.5, or 9.3) swell at pH 2.0, and this swelling may open cation-exchange sites that preferentially bind K+ to enable highly selective transport. The coated membranes also exhibit modest selectivity for K+ over H+, suggesting selective transport based on preferential partitioning of K+ into the coatings. Selectivity declines when increasing the source-phase KNO3 concentration to 0.1 M, perhaps because the discriminating transport pathway saturates. Moreover, selectivities are lower in electrodialysis than in Donnan dialysis, presumably because electrodialysis engages other transport mechanisms, such as electroosmosis and strong electromigration.

Dedoping of Lead Halide Perovskites Incorporating Monovalent Cations.

We report significant improvements in the optoelectronic properties of lead halide perovskites with the addition of monovalent ions with ionic radii close to Pb2+. We investigate the chemical distribution and electronic structure of solution processed CH3NH3PbI3 perovskite structures containing Na+, Cu+, and Ag+, which are lower valence metal ions than Pb2+ but have similar ionic radii. Synchrotron X-ray diffraction reveals a pronounced shift in the main perovskite peaks for the monovalent cation-based films, suggesting incorporation of these cations into the perovskite lattice as well as a preferential crystal growth in Ag+ containing perovskite structures. Furthermore, the synchrotron X-ray photoelectron measurements show a significant change in the valence band position for Cu- and Ag-doped films, although the perovskite bandgap remains the same, indicating a shift in the Fermi level position toward the middle of the bandgap. Such a shift infers that incorporation of these monovalent cations dedope the n-type perovskite films when formed without added cations. This dedoping effect leads to cleaner bandgaps as reflected by the lower energetic disorder in the monovalent cation-doped perovskite thin films as compared to pristine films. We also find that in contrast to Ag+ and Cu+, Na+ locates mainly at the grain boundaries and surfaces. Our theoretical calculations confirm the observed shifts in X-ray diffraction peaks and Fermi level as well as absence of intrabandgap states upon energetically favorable doping of perovskite lattice by the monovalent cations. We also model a significant change in the local structure, chemical bonding of metal-halide, and the electronic structure in the doped perovskites. In summary, our work highlights the local chemistry and influence of monovalent cation dopants on crystallization and the electronic structure in the doped perovskite thin films.

Efficient Synthesis of Ditopic Polyamide Receptors for Cooperative Ion Pair Recognition in Solution and Solid States.

Following a late-stage functionalization strategy, a series of heteroditopic cryptand receptors were prepared in three steps only from 1,4-dioxane. As evidenced by 1 H NMR spectroscopic and solid state analyses, these polyamide-crown ether conjugates present general ion pair binding capacity towards salts of monovalent cations and linear triatomic anions.

Duplex/quadruplex oligonucleotides: Role of the duplex domain in the stabilization of a new generation of highly effective anti-thrombin aptamers.

Recently, mixed duplex/quadruplex oligonucleotides have attracted great interest for use as biomedical aptamers. In the case of anti-thrombin aptamers, the addition of duplex-forming sequences to a G-quadruplex module identical or very similar to the best-known G-quadruplex of the Thrombin Binding Aptamer (HD1) results in new or improved biological properties, such as higher activity or different recognition properties with respect to HD1. Remarkably, this bimodular fold was hypothesized, based on its sequence, for the only anti-thrombin aptamer in advanced clinical trial, NU172. Whereas cation modulation of G-quadruplex conformation and stability is well characterized, only few data from similar analysis on duplex/quadruplex oligonucleotides exist. Here we have performed a characterization of structure and stability of four different duplex/quadruplex anti-thrombin aptamers, including NU172, in the presence of different cations and in physiological-mimicking conditions in comparison to HD1, by means of spectroscopic techniques (UV and circular dichroism) and differential scanning calorimetry. Our data show a strong reciprocal influence of each domain on the stability of the other and in particular suggest a stabilizing effect of the duplex region in the presence of solutions mimicking the physiological conditions, strengthening the idea that bimodular aptamers present better therapeutic potentialities than those containing a single G-quadruplex domain.

Effects of monovalent cations on folding kinetics of G-quadruplexes.

G-quadruplexes are special structures existing at the ends of human telomeres, the folding kinetics of which are essential for their functions, such as in the maintenance of genome stability and the protection of chromosome ends. In the present study, we investigated the folding kinetics of G-quadruplex in different monovalent cation environments and determined the detailed kinetic parameters for Na+- and K+-induced G-quadruplex folding, and for its structural transition from the basket-type Na+ form to the hybrid-type K+ form. More interestingly, although Li+ was often used in previous studies of G-quadruplex folding as a control ion supposed to have no effect, we have found that Li+ can actually influence the folding kinetics of both Na+- and K+-induced G-quadruplexes significantly and in different ways, by changing the folding fraction of Na+-induced G-quadruplexes and greatly increasing the folding rates of K+-induced G-quadruplexes. The present study may shed new light on the roles of monovalent cations in G-quadruplex folding and should be useful for further studies of the underlying folding mechanism.

Association of alkali metal cations with phosphatidylcholine liposomal membrane surface.

Interactions of alkali metal cations (Li+, Na+, K+, Cs+) with phosphatidylcholine (PC) liposomal membranes were investigated through experimental studies and theoretical considerations. Using a microelectrophoresis technique, charge densities of experimental membrane surfaces were measured as a function of the pH of electrolyte solutions. Equilibria between the PC liposomal membranes and monovalent ions were mathematically analyzed and described quantitatively through a previously proposed theoretical model. Association constants between functional groups of PC and the studied ions were determined and used to define theoretical curves of membrane surface charge density versus pH. Theoretical and experimental data were compared to verify the model. The PC membrane was found to have the highest affinity for lithium ions, among the ions tested.

The role of monovalent cations in the ATPase reaction of DNA gyrase.

Four new crystal structures of the ATPase domain of the GyrB subunit of Escherichia coli DNA gyrase have been determined. One of these, solved in the presence of K(+), is the highest resolution structure reported so far for this domain and, in conjunction with the three other structures, reveals new insights into the function of this domain. Evidence is provided for the existence of two monovalent cation-binding sites: site 1, which preferentially binds a K(+) ion that interacts directly with the α-phosphate of ATP, and site 2, which preferentially binds an Na(+) ion and the functional significance of which is not clear. The crystallographic data are corroborated by ATPase data, and the structures are compared with those of homologues to investigate the broader conservation of these sites.

Divalent cation-aided identification of physico-chemical properties of metal ions that stabilize RNA G-quadruplexes.

DNA and RNA sequences rich in guanosines (G) can form a four-stranded secondary structure known as a G-quadruplex (GQ), which plays a role in regulation of gene expression at the transcription and translation level. Both DNA and RNA GQs typically use the monovalent K(+) ion for stabilization of the structures. However, the fundamental reasons for K(+) acting as the most stabilizing metal ion for RNA GQs are not known. To identify the properties of a metal ion that stabilizes an RNA GQ we investigated the effect of alkaline earth metal cations and a set of divalent transition metal ions on two previously identified highly stable RNA GQs. Our results based upon circular dichroism and RNase T1 structure mapping data reveal that the RNA GQs are destabilized in the presence of the tested divalent metal cations. The destabilizing effect of a divalent metal cation is reversible upon increasing K(+) concentration. Results show that ionic radius, hydration energy, and binding strength towards the hard ligand (guanine O(6)) are important factors that determine a metal ion's ability to stabilize an RNA GQ. Additionally, the tested set of divalent metal cations incongruously affects RNA and DNA GQs.