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The production of ATP in mitochondria depends on the magnesium nuclear spin and magnetic moment of a Mg2+ ion in creatine kinase and ATPase. This suggests that enzymatic synthesis of ATP is an ion-radical process and thus depends on the external magnetic field (magnetobiology originates from this fact) and microwave fields, which control the spin states of ion-radical pairs and affect the ATP synthesis. The chemical mechanism of ATP synthesis and the origin of biological effects of electromagnetic (microwave) fields are discussed.

The interaction between the nicotinic acetylcholine receptor and Na+,K(+)-ATPase described previously was further studied in isolated rat diaphragm and in a membrane preparation of Torpedo californica electric organ. Three specific agonists of the nicotinic receptor: acetylcholine, nicotine and carbamylcholine (100 nmol/L each), all hyperpolarized the non-synaptic membranes of muscle fibers by up to 4 mV. Competitive antagonists of nicotinic acetylcholine receptor, d-tubocurarine (2 mcmol/L) or alpha-bungarotoxin (5 nmol/L) completely blocked the acetylcholine-induced hyperpolarization indicating that the effect requires binding of the agonists to their specific sites. The noncompetitive antagonist, proadifen (5 mcmol/L), exerted no effect on the amplitude of hyperpolarized but decreased K0.5 for this effect from 28.3 +/- 3.6 nmol/L to 7.1 +/- 2.3 nmol/L. Involvement of the Na+,K(+)-ATPase was suggested by data demonstrating that three specific Na+,K(+)-ATPase inhibitors: ouabain, digoxin or marinobufagenin (100 nmol/L each), all inhibit the hyperpolarizing effect of acetylcholine. Acetylcholine did not affectation either the catalytic activity of the Na+,K(+)-ATPase purified from sheep kidney or the transport activity of the Na+,K(+)-ATPase in the rat erythrocytes, i. e. in preparations not containing acetylcholine receptors. Hence, acetylcholine does not directly affect the Na+,K(+)-ATPase. In a Torpedo membrane preparation, ouabain (< or = 100 nmol/L) increased the binding of the fluorescent ligand: Dansyl-C6-choline (DCC). No ouabain effect was observed either when the agonist binding sites of the receptor were occupied by 2 mmol/L carbamylcholine, or in the absence Mg2+, when the binding of ouabain to the Na+,K(+)-ATPase is negligible. These results indicate that ouabain only affects specific DCC binding and only when bound to the Na+,K(+)-ATPase. The data obtained suggest that, in two different systems, the interaction between the nicotinic acetylcholine receptor and the Na+,K(+)-ATPase specifically involve the ligand binding sites of these two proteins.

A discovery of the huge magnesium isotope effect in enzymatic ATP synthesis provides a new insight into mechanochemistry of enzymes as the molecular machines. It has been found that the catalytic activity values of ATPase, creatine kinase and phosphoglycerate kinase are 2 to 4-fold higher once their active sites contain magnetic (25Mg) not spinless, non-magnetic (24Mg, 26Mg), magnesium cation isotopes. This clearly proves that the ATP synthesis is a spin-selective process involving Mg2+ as the electron accepting reagent. The formation of ATP takes place in an ion-radical pair resulted by two partners, ATP oxyradical and Mg+. The magnesium bivalent cation is a key player in this process, this ion transforms the protein molecule mechanics into a mere chemistry. This ion is a most critical detail of structure of the magnesium dependent phosphorylation enzymes as the mechanochemical molecular machines.

This review carries literary data on a biological effect, physiological and pathophysiological significance of magnesium ions for the organism. Diseases of hypo- and hypermagnesemia and mechanisms of regulation of magnesium content in the organism are reviewed in the article. There are cited data on the use of preparations, containing magnesium ions in pathologies of the kidneys, liver, gastrointestinal tract; in diabetes mellitus, arrhythmias, hypertensions, ischemias, myocardial infarction and other conditions. There are review data on the use of magnesium-containing mineral bishofit, preparations and balneologic agents on its basis.

Uterotonic peptide hormone oxytocin has been studied for its effect on ATP-dependent accumulation of Ca2+ nonsensitive to the effect of ruthenium red (10 microM) that is potentiated by Ca2+-precipitating anion - oxalate. That was studied in experiments made on the suspension of myometrium cells from estrogenized rats processed by digitonin (0.1 mg/ml) with the aim permeabilization of plasmatic membrane. The preliminary processing of intact myocytes by oxytocin solution (final concentration 100 mM) to permeabilization of plasmatic membrane causes partial inhibition (stimulated by oxalate by 25-30% (0-10 mM) of Mg2+, ATP-dependent accumulation of Ca2+ in nonmitochondrion intracellular calcium depot of perforates smooth-muscled cells. The value of the component of power-dependent accumulation of Ca2+, that is inhibited by oxytocin, increased with both the incubation time and oxalate concentration. Thapsigargin and cyclopiasonium acid, selective inhibitors of calcium pump of endo(sarco)plasmatic reticulum, when used in saturation concentration (50 mM and 10 mM, respectively), completely inhibit the component of Mg2+, ATP-dependent accumulation of Ca2+ in nonmitochondrial intracellular calcium depot that is stimulated with oxalate (< 10 mM) and inhibited with oxytocin (100 mM). It is concluded that oxytocin partially inhibits Mg2+, ATP-dependent calcium pump of myometrium cell endoplasmic reticulum.

The effect of sigetin (dipotassium salt, meso-3, 4-di-(n-sulfophenyl)- hexane) on the electrical and mechanical activity, Ca2+, Mg2+-ATPase activity and Mg2+, ATP-dependent Ca2+ transport in uterine smooth muscle have been investigated. Sigetin caused depolarization of the cell membrane, leading to Ca2+ dependent spike discharge and development of mechanical response. Sigetin also produced deceleration of the relaxation of carbachol contracture controlled mostly by the sarcolemmal ATP driven Ca2+ pump. The drug produced strong inhibition of Mg2+, ATP-dependent Ca2+ transport in sarcolemmal vesicles as well as the enzyme and transport activity of the solubilized Ca2+, Mg(2+)-ATPase reconstructed in liposomes. Under the experimental conditions close to the physiological (i.e. in the presence of an outwardly directed Ca2+ gradient) Ca2+ pump normally compensates passive release of this cation. Sigetin was found to shift this calcium equilibrium towards a non-compensated slow diffusion of the cation in the extravesicular space. Addition of sigetin to the incubation medium during the existence of the stationary transmembrane Ca2+ equilibrium stimulates transition of the stationary concentration of the cation in vesicles from high to lower level. The results suggest that the excitatory action of sigetin on myometrial smooth muscle may involve not only indirect stimulation of Ca2+ influx via the voltage operated Ca2+ channels but also through the inhibition of the ATP driven Ca2+ pump of the cell membrane.

The effect of ATP on bovine adrenal cortex pyruvate kinase has been studied. ATP is a competitive inhibitor of the enzyme, the Ki being 3.2 mM. Based on the efficiency of tryptophan fluorescence quenching, it was concluded that the magnesium complex of ATP is a true enzyme inhibitor. The role of Mg2+ in the inhibition process consists in the formation of a bridge between the enzyme and ATP. The ATP-dependent mechanism of pyruvate kinase inhibition is a potential physiological regulator of the enzyme determining the lower threshold of its sensitivity in vivo.

The effect of the membrane potential (K(+)-valinomycin system) on the Mg2+, ATP-dependent transport of Ca2+ in inside-out vesicles of myometrium sarcolemma has been studied. The membrane potential was identified by using a cyanine potential-sensitive probe, diS-C3-(5). In the presence of valinomycin (5.10(-8) M) the inside-out directed K+ gradient (delta psi = -86 mV, with a negative charge inside) stimulated the initial rate of the energy-dependent accumulation of Ca2+ transfer whereas the oppositely directed K+ gradient (delta psi = +72 mV, with a positive charge inside) had no effect on this process. The K+ gradient was formed by isotonic substitution of K+ in intra- or extravesicular space for choline +. At the same time, in the absence of K+ gradient the Mg2+, ATP-dependent accumulation of Ca2+ in membrane vesicles did not depend on the chemical nature of the cations (K+ or choline+) used for isotonicity. The decrease of delta psi from 0 to -86 mV affects the initial rate of Ca2+ accumulation but not the maximal content of the accumulated cation. Preliminary dissipation of the membrane potential (delta psi = -86 mV) in Mg2(+)-free isotonic (with respect of K+ and choline+) media containing ATP and Ca2+ resulted in the inhibition of Mg2+, ATP-dependent Ca2+ transport induced by subsequent addition of Mg2+. These results indicate that the negative (intravesicular) electrical potential activates the Ca-pump of smooth muscle sarcolemma. This activation is based on the increase in the turnover number of the Ca2+ transporting system but not on its affinity for the transfer substrate. The use of the absolute reaction rates theory made it possible to establish that the Ca-pump effectuates the transport of a single positive charge in inside-out vesicles of smooth muscle plasma membranes, i.e., the energy-dependent transport of Ca2+ occurs either as a symport (with an anion (Cl-) or an antiport with a monovalent cation (K+) or a proton. It is assumed that the potential dependence of the Ca-pump in the smooth muscle plasma membrane plays a role in the realization of effects of mediators and physiologically active substances that are manifested as stimulation of the contractile response and depolarization of the sarcolemma. In is quite probable that the delta psi-dependent Ca-pump is also responsible for the maintenance of intracellular homeostasis of monovalent cations (K+, H+, Cl-) in smooth muscle tissues.

The mechanism of substrate inhibition of rabbit skeletal muscle fructose-1.6-bisphosphatase was examined. Analysis of substrate saturation curves obtained at different concentrations of Mg2+ revealed that the inhibiting effect of the substrate is manifested only within the complex with Mg2+, whereas the free form of the substrate causes no inhibition. Evidence for the allosteric nature of substrate inhibition was obtained by partial desensitization of the enzyme in the presence of salicylates. It was shown that fructose-1.6-bisphosphatase inhibition by the substrate obeys positively cooperative kinetics and is noncompetitive with respect to the substrate involved in the catalytic process. Studies on enzyme modification in the presence of DTNB and pyridoxal-5'-phosphate demonstrated that the inhibiting concentrations of the substrate are bound to the center which differs from the allosteric site for AMP. It is suggested that the antagonism of simultaneous action of AMP and high substrate concentrations may be due to the competition of the phosphate groups of these ligands for binding to the common lysine residue located in the overlapping region of two allosteric sites.

In isolated smooth muscle of the rat portal vein removal of Mg2+ from perfusate induced rapid enhancement of frequency and amplitude of spontaneous contractions and decreased inhibiting effect of hypoxia on the smooth muscle contractile activity. Concomitant elevation of extracellular Ca2+ has no additional protective effect on the smooth muscle in hypoxia. Ten-fold lowering of Mg2+ content in buffer solution resulted in a significant rise in the tension of the smooth muscle activated by Ca2+. The data obtained suggest that protective effect of Mg2+-free solution on vascular smooth muscle in hypoxia may be mediated not only through increases intracellular Ca2+ concentration but also due to rising sensitivity of the smooth muscle cells contractile protein to Ca2+.

The growth of the hydrogen bacterium Hydrogenomonas eutropha Z-1 was studied in the conditions of continuous cultivation at different content of mineral elements in the medium. The physiological action of phosphorus, sulfur, potassium and magnesium in the bacterium was found to be displayed within a wide range of concentrations (mg/ml): P, 18-1400; S, 5.5-350; K, 4-550; Mg, 4.8-430. If the control of mineral elements was above or below these values, the specific growth rate of the cells decreased and their intracellular chemical composition changed.

The kinetic properties of creatinephosphokinase connected with the mytochondria of the heart were studied. The following kinetic parameters were measured: the Michaelis constants for all substrates -- MgATP, MgADP, creatine and creatinephosphate, and the maximal reaction rates for the direct and reverse reactions catalized by the mytochondrial iso-enzyme of creatinephosphokinase. The peculiarity of this iso-enzyme was demonstrated to consist in the independence of binding adeninenucleotides and guanidine substrates. The kinetic characteristics of this iso-enzyme are such that ATP synthesis from ADP and creatinephosphate is preferable: the Michaelis constant for MgADP is 0.05 mM, for creatinephosphate -- 0.5 mM, for MgATP -- 0.7 mM, for creatine -- 5.0 mM. It was also demonstrated that in the presence of all substrates and products of the reaction the ratio of the speeds of the direct and reverse reactions is regulated by Mg ions: with a low concentration of Mg (below that of ATP) the reaction of creatinephosphate synthesis takes place, with the Mg concentration growing the speed of ATP synthesis increases. Thus, an increase of Mg concentration can, depending on the conditions of the reaction, result in its complete reversal. The regulatory role of Mg is also demonstrated for the case in which ATP is synthesized in the reaction of oxidative phosphorylation, creatinephosphokinase-controlled reaction.

Calmodulin (CM) was found to activate adenylate cyclase (AC) in plasma membrane preparations of the rabbit heart in the presence of micromolar Ca2+ concentrations. CM action on the enzyme is terminated by trifluoperazine, troponin I and high Ma2 +/Ca2+ ratio. Isoproterenol in the presence of GTP, guanosine-5'-beta-gamma-imidotriphosphate[Cpp(NH)p] and sodium fluoride increases CM-dependent heart AC activity without changing the enzyme affinity to CM and Ca ions. CM has no effect on the convertion of regulatory N-protein into Gpp(NH)p-activated state. CM action on AC is revealed only if catalytic subunit forms a complex with regulatory N-protein. The conclusion was drawn that there is no distinct CM-dependent AC form in the heart, but the same catalytic subunit of the enzyme is regulated both by N-protein and CM. In the presence of Ca2+ and guanye nucleotides AC of the heart exists in the form of a complex: CM-catalytic subunit-N-protein.

The method of mathematical modelling and kynetic analysis was used to study the mechanism of the regulatory effect of magnesium ions on the creatinephosphokinase reaction in cardiac mitochondria. The regulatory effect of magnesium was shown to originate from the competition between ATP and ADP and the magnesium-complex-formation reaction, and the actual regulator proves to be MgADP. On the basis of the data obtained it is postulated that creatinephosphokinase in cardiac mitochondria is functionally connected with ATP-ADP-translocase. The obtained data permit to believe that the mitochondrial cratinephosphokinase is one of the key links in the process of energy transport in the cardiac cells.