Presence Of Bivalent Cations Research Articles

Cation Overcrowding Effect on the Oxygen Evolution Reaction.

The influence of electrolyte ions on the catalytic activity of electrode/electrolyte interfaces is a controversial topic for many electrocatalytic reactions. Herein, we focus on an effect that is usually neglected, namely, how the local reaction conditions are shaped by nonspecifically adsorbed cations. We scrutinize the oxygen evolution reaction (OER) at nickel (oxy)hydroxide catalysts, using a physicochemical model that integrates density functional theory calculations, a microkinetic submodel, and a mean-field submodel of the electric double layer. The aptness of the model is verified by comparison with experiments. The robustness of model-based insights against uncertainties and variations in model parameters is examined, with a sensitivity analysis using Monto Carlo simulations. We interpret the decrease in OER activity with the increasing effective size of electrolyte cations as a consequence of cation overcrowding near the negatively charged electrode surface. The same reasoning could explain why the OER activity increases with solution pH on the RHE scale and why the OER activity decreases in the presence of bivalent cations. Overall, this work stresses the importance of correctly accounting for local reaction conditions in electrocatalytic reactions to obtain an accurate picture of factors that determine the electrode activity.

Simultaneously Antimicrobial, Protein-repellent and Cell-compatible Polyzwitterion Networks: More Insight on Bioactivity and Physical Properties.

A poly(oxanorbornene)-based polyzwitterion with primary ammonium and carboxylate groups (PZI) has been reported previously as the first simultaneously antimicrobial and protein-repellent polyzwitterion. Here, additional physical and biological properties of three poly(oxanorbornene)-based polyzwitterions with different functional groups (PZI, the polycarboxybetaine PCB, and the polysulfobetaine PSB) are compared to understand the molecular origins of this unusual bioactivity. Additionally, the three polyzwitterions and the antimicrobial, polycationic SMAMP are exposed to proteins, bacteria suspensions, human plasma and serum. These interactions are investigated by surface plasmon resonance spectroscopy. In protein adhesion studies, neither fibrinogen nor lysozyme adhere irreversibly to PZI, yet reversible interaction with lysozyme is observed at pH 7 and 8. In the presence of bivalent cations, reversible fibrinogen adhesion on PZI and PSB is observed, but not on PCB. This might explain why mammalian cells grow on PZI and PSB, but not on PCB. PZI does not show human plasma adhesion, while PCB and PSB have 0.27 and 0.48 ng mm-2 adhered plasma, and SMAMP even 6.3 ng mm-2. Both PZI and SMAMP show strong serum adhesion, while no serum adhered to PCB, and only little to PSB. This could be related to the pH difference between serum and plasma, to which the pH-responsive primary ammonium groups are susceptible, while the permanently charged NR4 + groups are unaffected. Both PZI and PCB showed none or only little bacterial adhesion. PCB is also intrinsically antimicrobial against E. coli and S. aureus bacteria and thus is also simultaneously protein-repellent and antimicrobially active. Thus, while the carboxylate groups of PZI and PCB seems to be a prerequisite for the dual antimicrobial activity and protein-repellency, the pH-responsiveness of the primary ammonium group seems to make the PZI molecule vulnerable for protein adhesion in fluids that are slightly out of the physiological range.

Acid-induced gelation of enzymatically cross-linked caseinate in different ionic milieus

Acid casein powder was used to prepare caseinate solutions (27 g/kg) with different ionic milieus: sodium caseinate (NaCN, I ∼0.015 mol/L) and calcium caseinate (CaCN, I ∼0.03 mol/L) were obtained by neutralisation with NaOH and Ca(OH)2, respectively, and dissolving in phosphate buffer resulted in a high ionic strength caseinate solution (CN-PB, I ∼0.16 mol/L). Treatment with microbial transglutaminase (mTGase) for defined incubation times lead to different extents of casein cross-linking, which were characterised by size exclusion chromatography and the N-ε-(γ-glutamyl)-lysine isopeptide content (IC). Maximum polymer size was reached at ∼90% casein polymerisation, and increased in the order NaCN < CN-PB < CaCN. Further enzyme treatment, however, increased the IC, pointing to cross-links within existing polymers. We suggest that the maximum polymer size is determined by the size of casein particles resulting from self-assembly in solution and that mTGase preferably acts on molecules within the same particle. Enhanced association at higher ionic strength (CN-PB) or in the presence of bivalent cations (CaCN) may therefore result in larger covalently cross-linked casein polymers. Furthermore, oscillation rheometry revealed that the relationship between casein cross-linking and stiffness of gels acidified with glucono-δ-lactone depends on the ionic milieu. While for NaCN G'MAX increased with the time of cross-linking, the presence of ions resulted in the highest G'MAX at moderate cross-linking intensities. This was also observed when NaCl was added to cross-linked NaCN. The results suggest that electrostatic attraction during gel formation are interfered by ions and cannot be compensated by rearrangements in case of extensively cross-linked casein particles.

Fe(III) nucleation in the presence of bivalent cations and oxyanions leads to subnanoscale 7 Å polymers.

Highly disordered Fe(III) phases formed in the presence of bivalent cations and oxyanions represent important components of the global Fe cycle due to their potential for rapid turnover and their critical roles in controlling the speciation of major and trace elements. However, a poor understanding of the formation pathway and structure of these Fe phases has prevented assessments of their thermodynamic properties and biogeochemical reactivity. In this work, we derive structural models for the Fe(III)-As(V)-Ca and Fe(III)-P-Ca polymers formed from Fe(II) oxidation and Fe(III) polymerization in the presence of As(V)/P and Ca. The polymer phase consists of a less than 7 Å coherent network of As(V)/P coordinated to Fe(III) polyhedra, with varying amounts of Ca bound directly and indirectly to the oxyanion. This phase forms at the onset of Fe(II) oxidation and, because of its large oxyanion:Fe solids ratio, depletes the oxyanion concentration with only small amounts of Fe. Our results demonstrate that when a steady supply of Fe(III) is provided from an Fe(II) source, these Fe(III) polymers, which dominate oxyanion uptake, form with little dependence on the initial oxyanion concentration. The formation mechanisms and structures of the oxyanion-rich Fe(III) polymers determined in this study enable future thermodynamic investigations of these phases, which are required to model the interrelated biogeochemical cycles of Fe, As(V)/P, and Ca.

The effect of some mono and bivalent metal cations on the individual hydrogen bond energies in A−T and G−C base pairs

The effect of interactions of various Ia and IIa cations with two positions of the adenine-thymine (A-T) and guanine-cytosine (G-C) base pairs on the geometries and individual hydrogen bond (HB) energies have been investigated by using the atoms in molecules (AIM) method at the B3LYP/6-311++G(d,p) level of theory. The cations that possess higher charge/radius (q/rad) ratio make higher changes on the individual HB energies and the total binding energies of G-C base pair, while no meaningful changes are observed for the A-T base pair. The effect of coordination with cations on the H-bond energies of A-T type complexes, especially in the X'-A-T derivatives, is slight and different from the GC complexes. The changes of total binding energies are most important in the presence of bivalent cations. The estimated individual EHBs are in good agreement with the results of natural bond orbital (NBO) analysis and also with the structural parameters of HBs.

Structure and activity of the only human RNase T2

Mutations in the gene of human RNase T2 are associated with white matter disease of the human brain. Although brain abnormalities (bilateral temporal lobe cysts and multifocal white matter lesions) and clinical symptoms (psychomotor impairments, spasticity and epilepsy) are well characterized, the pathomechanism of RNase T2 deficiency remains unclear. RNase T2 is the only member of the Rh/T2/S family of acidic hydrolases in humans. In recent years, new functions such as tumor suppressing properties of RNase T2 have been reported that are independent of its catalytic activity. We determined the X-ray structure of human RNase T2 at 1.6 Å resolution. The α+β core fold shows high similarity to those of known T2 RNase structures from plants, while, in contrast, the external loop regions show distinct structural differences. The catalytic features of RNase T2 in presence of bivalent cations were analyzed and the structural consequences of known clinical mutations were investigated. Our data provide further insight into the function of human RNase T2 and may prove useful in understanding its mode of action independent of its enzymatic activity.

Activity and properties of fructose bisphosphatase of turbellaria Phagocata sibirica

Activity and properties of fructose bisphosphatase (FBPase) was studied in the free-living turbellarian Phagocata sibirica. All subcellular fractions of P. sibirica (12 000 g cytosol, 105 000 g cytosol, mitochondria, and microsomes) have the FBPase activity. There was studied dependence of the FBPase reaction rate on the substrate concentration. For realization of the enzyme activity, the high affinity to substrate and presence of bivalent cations (Mg2+ or Mn2+) are necessary. The was studied the effect of various effectors as well as of monovalent (Na+, K+, Li+, and NH4+) and bivalent (Zn2+ and Cu2+) cations.

Glycosyl conjugates of biotinylated diaminopyridine applied for study of carbohydrate-to-carbohydrate interaction

Previous studies by us and others established that cell-cell adhesion is mediated by specific carbohydrate-to-carbohydrate interaction (CCI). Those previous studies were based on various biochemical and biophysical approaches, including the use of labeled glycosyl epitopes with fluorescent tag. However, these methods ideally require that the glycosyl epitope must be fixed to a solid phase molecule, preferably with multivalency. The purpose of the present study is to establish a CCI process using specific glycosyl residues conjugated to biotinylated diaminopyridine (BAP), and to observe: (i) clear occurrence of homotypic CCI between "Os Fr.B" having 5-6 GlcNAc termini, vs. absence of such homotypic CCI between "Os Fr.1" having 2 GlcNAc termini; (ii) occurrence of heterotypic CCI between GM3 ganglioside and Os Fr.B, vs. absence of such heterotypic CCI between GM3 and Os Fr.1. Interaction between Os Fr.B-BAP conjugate and Os Fr.B-ceramide mimetic (Os Fr.B-mCer) was demonstrated based on two experiments: (i) dose-dependent binding of Os Fr.B-BAP conjugate to polystyrene plates coated with Os Fr.B-mCer was observed in the presence of bivalent cation, a prerequisite for all CCI processes, and such binding was abolished by EDTA; (ii) binding between equal nanomolar Os Fr.B-BAP and Os Fr.B-mCer was inhibited by mM concentration Os Fr.B without conjugate, in dose-dependent manner. Thus, cell adhesion processes based on homotypic CCI between N-linked glycans having multiple GlcNAc termini, and heterotypic CCI between GM3 and such glycans, were clearly observed using BAP conjugates of glycosyl epitopes.

Fructose bisphosphatase of cestodes of Bothriocephalus scorpii

There have been studied the activity and properties of fructose bisphosphatase (FBPase) of Bothriocephalus scorpii parasitizing in pyloric appendages of the goby Myoxocephalus brandti. All subcellular fractions of B. scorpii (12 g/cytosol, 105 g/cytosol, mitochondria, and microsomes) have the FBPase activity. The enzyme has a high affinity to substrate and needs the presence of bivalent cations (Mg2+ or Mn2+). AMP inhibits the enzyme significantly. Action of various effectors has been studied. The monovalent (Na+, K+, Li+, NH4+) and bivalent cations (Zn2+ and Cu2+) inhibit the enzyme activity.

Clay mineral-enhanced bioremediation of marine oil pollution

Three year old laboratory mixtures of Prestige oil, Atlantic sea water and various types of clay mineral powder (Na- and Ca-smectites, palygorskite, synthetic mica-montmorillonite and a low defect kaolinite) contained significantly lower concentrations of extracted oil than the non-clay containing control. As no significant quantities of oil could be detected in the clay samples following solvent extraction, and no expansion of the viscous oil-treated smectite interlayers could be detected by X-ray diffraction, the measured hydrocarbon content is suggested to provide a reasonable estimate of the total oil present. The observed decrease in the concentration of oil over the reaction period is attributed largely to bacterial digestion rather than to hydrocarbon adsorption, with denser populations of cultured oleophilic bacteria in the more strongly digested samples. Overall, the smectites with high specific surface areas and cation exchange capacities induced the highest degree of oil breakdown, and also stimulated the digestion of the heavier aromatic and resin–asphaltene compounds. The presence of bivalent cations at interlayer sites and hydrated surfaces, including abundant Mg 2+ derived from sea water, is suggested to minimize the thickness of the diffuse ion layer and thus enhance both inorganic and organic nutrient supply from mineral surface to bacterial cell. Adding clay minerals to marine oil pollution is therefore suggested as an effective and economically interesting way of enhancing the bacterial digestion of contaminating spills.

The role of extracellular calcium in exo- and endocytosis of synaptic vesicles at the frog motor nerve terminals

In the present study we combined FM 1-43 imaging and electrophysiological recording of miniature end-plate currents (MEPCs) to determine the role of extracellular calcium in synaptic vesicle exo- and endocytosis at the frog motor nerve terminals. We replaced extracellular Ca 2+ ions with other bivalent cations (Sr 2+, Ba 2+, Cd 2+, Mg 2+) or used a calcium-free solution and monitored fluorescent staining of the nerve terminals in the presence of caffeine, which promotes the release of Ca 2+ from intracellular stores. Caffeine has induced FM1-43 internalization only in the presence of bivalent cations in the external solution. The exposure of the neuromuscular junction to caffeine in a calcium-free solution caused a reversible failure of FM 1-43 loading and an increase in the nerve terminal width. This effect of a calcium-free solution was not due to a decrease in exocytosis, because caffeine-induced FM1-43 unloading from the previously loaded nerve terminals, as well as a degree of the MEPCs frequency increase, was unchanged. We conclude that the presence of Ca 2+ or other bivalent cations in extracellular space is necessary for endocytosis but not for exocytosis of synaptic vesicles, while transmitter release is promoted by efflux of Ca 2+ from intracellular stores. The effect of extracellular Ca 2+ on endocytosis might be driven by the non-specific interactions with membrane lipids.

Biophysical characterization of S100A8 and S100A9 in the absence and presence of bivalent cations

S100A8 and S100A9 are two proinflammatory molecules belonging to the S100 family of calcium-binding proteins. Common to all S100 proteins S100A8 and S100A9 form non-covalently associated complexes which have been shown to exhibit different functional properties. Besides dimerization, recent research is focused on the importance of higher oligomeric structures of S100 proteins induced by bivalent cations. While S100A8/S100A9-heterodimers are formed in the absence of calcium, tetramerization is strictly calcium-dependent. Heterodimer formation is not a simple process and our biophysical analyses (FRET, ESI-MS) demonstrate that simply mixing both subunits is not sufficient to induce complex formation. Steps of denaturation/renaturation are necessary for the recombinant complex to show identical biophysical properties as S100A8/S100A9 obtained from granulocytes. In addition to calcium both proteins are able to bind zinc with high affinity. Here we demonstrate for the first time by different biophysical methods (MALDI-MS, ESI-MS, fluorescence spectroscopy) that zinc-binding, like calcium, induces (S100A8/S100A9) 2-tetramers. Using mass spectrometric investigations we demonstrate that zinc triggers the formation of (S100A8/S100A9) 2-tetramers by zinc-specific binding sites rather than by interactions with calcium-specific EF-hands. The zinc-induced tetramer is structurally very similar to the calcium-induced tetramer. Thus, like calcium, zinc acts as a regulatory factor in S100A8/S100A9-dependent signaling pathways.

Monitoring of integrin-mediated adhesion of human ovarian cancer cells to model protein surfaces by quartz crystal resonators: evaluation in the impedance analysis mode

The quartz crystal microbalance (QCM) was used to monitor specific, integrin-mediated adhesion of human ovarian cancer cells to distinct extracellular matrix (ECM) proteins immobilized on gold-coated quartz crystal resonators. The QCM was operated in the impedance analysis mode, where frequency shift as well as bandwidth are accessible on a broad range of overtones. The increase in bandwidth caused by covering the quartz resonator with cells was reproducible and largely independent of overtone order, whereas the frequency shift displayed some variability. Thus the bandwidth proved to be the more robust parameter for sensing cell adhesive events. The bandwidth increased in proportion to the number of seeded cells to the quartz crystal as long as the number was below 150,000 cells/ml. Comparing the resonance parameters on different harmonics, one finds that viscoelastic modeling with homogeneous layer systems cannot reproduce the results: lateral heterogeneity has to be taken into account. The differences in adhesive strength of human ovarian cancer cells towards selected ECM proteins monitored by QCM was in good agreement with data obtained by conventional cell adhesion assays. Strong cell adhesion was observed to the ECM proteins vitronectin (VN) and fibronectin (FN), while only weak attachment occurred on laminin. In order to prove specific, integrin αvβ3-mediated cell adhesion to its ligands FN and VN, the cyclic integrin αvβ3-directed peptide c(RGDfV) was used as competitor and significantly reversed cell adhesion. Since integrin interaction with ECM proteins is dependent on the presence of bivalent cations, cell detachment was also seen after treatment of cell monolayers with the chelator ethylene-dinitro-tetra-acetic acid (EDTA). The QCM technique is a reliable method to monitor cell adsorption to ECM-pretreated surfaces in real time. It may be an alternative tool for screening specific and selective antagonists of integrin/ECM interaction.

Characteristics of Single Ionic Channels Induced by Sulfonic Acid Derivatives of Dibenzo-18-Crown-6 in Bilayers

Sulfonic acid derivatives of dibenzo-18-crown-6 (DB18C6) have been shown to induce formation of ion channels permeable to monovalent cations in bilayer lipid membranes (BLM). Some electric characteristics of channels have been studied by means of the voltage clamp method. Channels displayed little interionic discrimination. The voltage dependence as well as the multiple state behaviors of the channels functioning was observed. The channels formation has been shown to be dependent on pH of bath solutions and on the presence of bivalent cations in them. The channels supposed to be formed from aggregates of complexes associates assembled in aqueous salt solutions.

Fatty acids enhance membrane permeabilization by pro-apoptotic Bax.

Fatty acids are known promoters of apoptosis. In the present study, the direct role of fatty acids with regard to their ability to cause membrane permeabilization by Bax was explored. Addition of fatty acids to liposomes in the presence of cations greatly enhanced the permeabilizing activity of Bax, a pro-apoptotic Bcl-2 protein. This provides a putative mechanism for the role of fatty acids in apoptosis. It is not a result of detergent-like properties of fatty acids, since a different micelle-forming amphiphile, dilysocardiolipin, was strongly inhibitory. We also demonstrate that there is a synergistic effect on Bax-induced permeabilization between Ca(2+) and Mg(2+), both on the binding of Bax to liposomes as well as on the induction of the leakage of liposomal contents. Micromolar concentrations of Ca(2+) added externally or submicromolar concentrations of free Ca(2+) present in the medium were sufficient to promote Bax-induced permeabilization synergistically with externally added Mg(2+). These results indicate that Bax can induce leakage from liposomes at ion concentrations resembling those found physiologically. The synergistic effects of Ca(2+) and Mg(2+) were observed with liposomes with different lipid compositions. Thus the action of Bax is strongly modulated by the presence of bivalent cations that can act synergistically, as well as by micelle-forming lipid components that can be either stimulatory or inhibitory.

Phase separation in calcium alginate gels.

Alginates are polysaccharides consisting of beta-D-mannuronate and alpha-L-guluronate units. In the presence of bivalent cations like calcium the guluronate blocks form physically cross-linked gels. The gelation properties of alginates play an important role in the stability of extracellular polymer substances and in the food industry. When stock solutions of Ca2+ ions and alginate are mixed, the gelation starts before the Ca2+ ions are evenly distributed, which leads to non-uniform gels. In this contribution, Ca alginate gels were prepared by in situ gelation using glucono-delta-lactone and CaCO3. In this way, uniform gels could be prepared directly in the measuring cell. Below a critical concentration, highly viscous solutions were obtained, which were below the critical point of gel formation. In these solutions at low rotational speeds a Schlieren peak arose, which became smaller and steeper with increasing time until a new meniscus could be detected. This behaviour is in contrast to the peak broadening due to diffusion after a synthetic boundary was formed. Evaluation of the data leads to negative diffusion coefficients. It has been shown by others that the mutual diffusion coefficient must be negative in the spinodal region. This phenomena is known as uphill diffusion and leads to phase separation of a binary system. The formation of the gel phase in this case is therefore discussed as uphill diffusion.

Virus retention and transport through Al-oxide coated sand columns: effects of ionic strength and composition

Knowledge of the factors that influence the fate and transport of viruses in porous media is very important for accurately determining groundwater vulnerability and for developing protective regulations. In this study, six saturated sand column experiments were performed to examine the effects of a positively charged Al-oxide, which was coated on sand particles, on the retention and transport of viruses (φX174 and MS-2) in background solutions of different ionic strength and composition. We found that the Al-oxide coating on sand significantly removed viruses during their transport in a phosphate buffered saline (PBS) solution. Mass balance calculations showed that 34% of the input MS-2 was inactivated/irreversibly sorbed on the surface of Al-oxide coated sand whereas 100% of φX174 was recovered. Results from this study also indicated that higher ionic strength facilitated the transport of both φX174 and MS-2 through the Al-oxide coated sand. This was attributed to the effect of ion shielding, which at higher ionic strength decreased the electrostatic attraction between the viral particles and the sand surface and consequently decreased virus sorption. Strong effect of the ionic strength indicates that an outer-sphere complexation mechanism was responsible for the virus sorption on the Al-oxide coated sand. Ion composition of the background solutions was also found to be a significant factor in influencing virus retention and transport. Virus transport was enhanced in the presence of phosphate (HPO 4 2−) as compared to bicarbonate (HCO 3 −), and the effect of HPO 4 2− was more significant on MS-2 than on φX174. The presence of bivalent cations (Ca 2+ and Mg 2+) increased virus transport because the cations partially screened the negative charges on the viruses therefore decreased the electrostatic attraction between the positively charged sand surface and the negatively charged viruses. Mass recovery data indicated that bivalent cations gave rise to a certain degree of inactivation/irreversibly sorption of φX174 on the surface of Al-oxide coated sand. On the contrary, the bivalent cations appeared to have protected MS-2 from inactivation/irreversibly sorption. This study provides some insights into the mechanisms responsible for virus retention and transport in porous media.

Removal of arsenic from groundwater using low cost ferruginous manganese ore

A low cost ferruginous manganese ore (FMO) has been studied for the removal of arsenic from groundwater. The major mineral phases present in the FMO are pyrolusite and goethite. The studied FMO can adsorb both AS(III) and As(V) without any pre-treatment, adsorption of As(III) being stronger than that of As(V). Both As(III) and As(V) are adsorbed by the FMO in the pH range of 2–8. Once adsorbed, arsenic does not get desorbed even on varying the pH in the range of 2–8. Presence of bivalent cations, namely, Ni 2+, Co 2+, Mg 2+ enhances the adsorption capability of the FMO. The FMO has been successfully used for the removal of arsenic from six real groundwater samples containing arsenic in the range of 0.04–0.18 ppm. Arsenic removals are almost 100% in all the cases. The cost of the FMO is about 50–56 US$ per metric tonne.

Guanosine nucleotides regulate B2 kinin receptor affinity of agonists but not of antagonists: discussion of a model proposing receptor precoupling to G protein.

The effect of nucleotides on binding of the B2 kinin (BK) receptor agonist [3H]BK and the antagonist [3H]NPC17731 to particulate fractions of human foreskin fibroblasts was studied. At 0 degrees C, particulate fractions exhibited a single class of binding sites with a Kd of 2.3 nM for [3H]BK and a Kd of 3.8 nM for the antagonist [3H]NPC17731. Incubation with radioligands at 37 degrees C for 5 min gave a reduction of agonist, as well as antagonist, binding that was between 0-40% depending on the preparation, even in the absence of guanosine nucleotides. As shown by Scatchard analysis, this reduction in specific binding was due to a shift in the affinity of at least a fraction of the receptors. The presence at 37 degrees C of the guanine nucleotides GTP, GDP and their poorly hydrolyzable analogs left [3H]NPC17731 binding unaffected, but reduced the receptor affinity for [3H]BK to a Kd of about 15 nM. The maximal number of receptors, however, was unchanged. This affinity change was strongly dependent on the presence of bivalent cations, in particular Mg2+. It was reversed by incubation at 0 degrees C. The rank order of the guanosine nucleotides for [3H]BK binding reduction was GTP[gammaS] = Gpp[NH]p > GTP = GDP > GDP[betaS]. GMP, ATP, ADP and AMP showed no influence on agonist binding. A model for the interaction of the B2 kinin receptor with G proteins is discussed.

The access of metabolites into yeast mitochondria in the presence and absence of the voltage dependent anion selective channel (YVDAC1).

Since yeast Saccharomyces cerevisiae mutants depleted of the voltage dependent anion selective channel (YVDAC1) are still able to grow on a non-fermentable carbon source, a functional transport system in the outer mitochondrial membrane must exist to support the access of metabolites into mitochondria. It was assumed that the properties of the system could be inferred from the differences in the results observed between wild type and mutant mitochondria since no crucial differences in this respect between the two types of mitoplasts were observed. YVDAC1-depleted mitochondria displayed a highly reduced permeability of the outer membrane, which was reflected in increased values of K0.5(NADH) for respiration and K0.5(ADP) for triggering phosphorylating state as well as in delayed action of carboxyatractylate (CATR) in inhibition of phosphorylating state. The parameters were chosen to express the accessibility of the applied species to the intermembrane space. The passage of the molecules through the outer membrane depleted of YVDAC1 could be partially improved in the presence of bivalent cations (Mg2+, Ca2+), as in their presence lower values of the calculated parameters were obtained. The restrictions imposed on the transport of molecules through the YVDAC1-depleted outer membrane resulted in a competition between them for the access to the intermembrane space as measured by changes in parameters observed for a given species in the presence of another one. The competition was stronger in the absence of Mg2+ and depended on charge and size of transported molecules, as the strongest competitor was CATR and the weakest one--NADH. Thus, it can be concluded that the transport system functioning in the absence of YVDAC1 is modulated by bivalent cations and charge as well as size of transported molecules. Since an increased level of respiration due to the dissipation of delta psi causes an increase of K0.5(NADH) in both wild type and YVDAC1-depleted mitochondria it is concluded that a common property of YVDAC1 and the system functioning in YVDAC1-depleted mitochondria seems to be the dependence of the capacity on the level of mitochondrial respiration.