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The relationship between metabolism of the main myocardial amino acids, glutamate, aspartate and alanine, and energy state of hypoxic myocardium, was studied. Depression of cardiac contractile function during asphyxia in rats was accompanied by a decrease in glutamate mitochondrial and tissue contents and an increase in the tissue alanine and mitochondrial aspartate. The reduction of mitochondrial glutamate in asphyxia was related to losses of intramitochondrial ATP and state 3 respiration with glutamate and malate. Using NMR technique, exogenous glutamate and oxaloacetate were shown to increase succinate formation coupled with ATP and CTP production in the rat heart mitochondria in absence of aeration. These data suggest that glutamate and products of its transamination decrease the contraction of hypoxic myocardium stimulating anaerobic energy formation in the tricarboxylic acid cycle.

The state I in equilibrium state II transients in bean leaves were studied by ESR method at various temperatures. P700 photooxidation induced by far-red light after leaf adaptation to darkness demonstrated two phases. This kinetics correlated with two overshuts in the kinetics of P700+ reduction observed after additional switching on the light exciting Photosystem II. In the atmosphere of gaseous nitrogen P700 behaviour was reversibly modified. Multiphase character of P700 kinetics correlated with the state I in equilibrium state II phenomenon as measured by chlorophyll a fluorescence from Photosystem II. The rate-limiting step at the acceptor side of Photosystem I electron transport chain may be considered as a point controlling the state of intact chloroplasts.

The free radical EPR signals of ubisemiquinone in mitochondria and submitochondrial particles (SMP) were investigated. One of the signals observed under the conditions of the respiratory chain highly oxidized and characterized by an unusually short time of the spin-lattice relaxation has previously been termed as SQ-2. The intensity of SQ-2 in SMP strongly depends on pH, the maximal concentration of QH. is reached at about 8.5. The signal is absent in the succinate dehydrogenase-depleted SMP and is highly sensitive to specific inhibitors of succinate: CoQ-oxidoreductase, such as alpha-thenoyltrifluoroacetone and carboxin. In SMP SQ-2 disappears in the presence of low concentrations of ferricyanide, while in mitochondria this non-penetrating oxidant provokes the appearance of SQ-2. The data obtained suggest that SQ-2 belongs to a stable ubisemiquinone which forms a complex with a FeS center of succinate dehydrogenase, is localized at the M-side of the membrane, and is kinetically isolated from the cytochrome chain. Oxidation of the terminal segment of the respiratory chain of mitochondria and SMP reduced by succinate in the presence of antimycin, is in some cases accompanied by an appearance of a strong free radical EPR signal which is stable at 77K but disappears rapidly in the frozen samples at -30- -40 degrees C. It is suggested that the signal is generated by an antimycin-insensitive oxidation of QH2 to QH. via the branch of the respiratory chain comprised of the Rieske FeS-protein and cytochrome c1. The mechanisms of how the two-electron oxidation-reduction of CoQ is coupled with the one-electron transfer through the cytochromes and FeS centers in the respiratory chain are discussed.

A study of interaction between ten different spin labels and bean chloroplast membranes was carried out. It was found that spin labels were localized in three characteristic regions of the membrane. These regions differ in density and mode of the lipids package. The processes of light--induced reduction (with loss of paramagnetism) and reoxidation of spin labels are due to the interaction with the components of electron-transport chain between photosystems I and II. The kinetics of redox-changes of spin labels depends on the site of the paramagnetic fragment location in membrane.

3-Phosphono-2-imino-1-methyl-4-oxoimidazolidine (PIMOI), AMP and p-nitrophenyl phosphate (pNPP) were dephosphorylated in the presence of rat heart cytosol at 37 degrees C pH 6.3 at the rates of 0.71, 0.45 and 1.07 mumol/mg X h, respectively. When mixed together, these compounds inhibited the hydrolysis of each other, which points to the participation of common enzyme(s) in this process. The inhibitor of 5'-nucleotidase (alpha,beta-methylene)-ADP, did not affect PIMOI cleavage and moderately inhibited AMP hydrolysis (by ADP, did not affect PIMOI cleavage and moderately inhibited AMP hydrolysis (by 30-50%), thus suggesting that acidic phosphatases are responsible for PIMOI and AMP hydrolysis under these conditions (pH 6.3). Phosphocreatine (PCr) and phosphocyclocreatine (PcCr) were stable to hydrolysis by the cytosolic fraction. However, addition of AMP to the medium containing PCr or PcCr resulted in AMP phosphorylation down to ATP due to the effects of these phosphagens and, probably, of microcontaminations of ATP. This was followed by gradual disappearance of PCr or PcCr and by accumulation of Pi as a result of the "ATPase" activity in the cytosol. The hydrolysis of AMP, PIMOI and p-NPP was sensitive to sulfhydryl reagents [5,5'-dithio-bis-(2-nitrobenzoate) and, in part, 2,4-dinitro-fluorobenzene] and fluoride ion. Thus, PIMOI is a competitive substrate of acidic phosphatases in heart cytosol with respect to AMP and p-NPP. This may partly explain the protective effect of PIMOI on ischemic myocardium.

The possibility of in vitro lysis of fibrinous clots was shown experimentally (a model of blood flow in a vessel incompletely occluded by a thrombus) using the magnet-directed transport of trypsin immobilized on a magnet-susceptible carrier. During the clot lysis no fibrinolytics were recorded in the solution washing the clots, and lysis took place under the influence of trypsin on the magnetic carrier which precipitated on the surface of the clot and remained there for the necessary time (about 2 hours). The method of thrombus lysis suggested allows the dose of the injected drug to be decreased and does not require therapeutic concentration of thrombolytics in the general blood flow.

The parameters of fluorescence spectra of myosin and its subunits in Tris-HCl-buffer (pH 7.2) were studied. Analysis of the experimental results of myosin fluorescence quenching with I-ions and the quantum yield of the fluorescence at the excitation wavelength 296 nm shows that the greater part of the tryptophan residues (21 out of 28) is located in the hydrophylic environment. Concentrated solutions of NaCl and KCl do not affect myosin fluorescence, while LiCl, which changes the quaternary structure of the protein, brings about a change in the parameters of the myosin fluorescence spectra. This may be linked with structural changes accompanying the dissociation of the ligh subunits of myosin in the presence of LiCl.

The kinetics P700 redox transients in bean chloroplasts induced by short (t 1/2 = 7 microsecond) flashes of white light has been studied. The flashes were fired in the dark or simultaneously with the continuous background far red light (707 nm). After oxidation of electron transport chain by 707 nm light only 50% of P700+ centres were reduced by photosystem 2 in response to the first flash of saturating intensity. The total reduction of P700+ centres was observed only after two or more saturating flashes. The similar data were obtained for various native photosynthetic systems (bean leaves and leaves of other higher plants). It has been shown that the quantitative difference in P700 behaviour after the first and following flashes may be interpreted in terms of a mechanism of the plastoquinol oxidation by photosystem 1. Twofold oxidation of one molecule of plastoquinol was assumed as its successive oxidation to plastosemiquinone and plastoquinone by different acceptors localized nea the same P700 centre (cytochrome f and plastocyanine). It has been shown that one molecule of plastoquinol was arised due to the interaction of two plastosemiquinones reduced by different P680 centre.

The interrelated temperature induced structural changes in the protein and lipid fractions of the Na+, K+-ATPase preparations were studied. A spin-labelled analog of ATP (ATP) was used, where the paramagnetic fragment was attached to the 2' (3')-OH ribose group. It is shown that the rotational mobility of ATP changes with the temperature in a discontinuous way. This correlates with the behaviour of the enzymatic activity of Na+, K+-ATPase and with the state of lipids in the enzyme preparations, both being characterized by a break in the 20-23 degrees C temperature region. When the Mn2+ ions were substituted by Mn2+ a strong magnetic dipolar interaction between the spin label in ATP and the Mn2+ ion was observed, which proves that the complex E-ATP--Mn2+ is formed before the hydrolysis of ATP. The structural changes which occur near 20 degrees C were also observed in the neighbourhood of the SH-groups modified by spin label X, an analog of maleimide. The structural changes observed support the idea that the protein-lipid interactions in the Na+, K+-ATPase provide the relaxation of the system from the unstable state during hydrolysis of ATP and cation transport.

The work was undertaken for the purpose of studying the interaction of human blood plasma lipoproteins (LP) with bi-layer lipid membranes (BLM), which is regarded to be a model of nonreceptive interaction of LP with the cell membranes. The surface LP of very low density (LPVLD) layer is much "denser" in hyperalpha-lipoproteinemia than in hypoalpha-lipoproteinemia. It was found that atherogenic LP classes induce an increase in BLM conductivity in much lesser concentration than the subclasses of antiatherogenic LP of high density. LPVLD of individuals with their increased content in plasma increase the electrical capacity of BLM to a lesser extent than LPVLD of healthy individuals. In most experiments with LPVLD of individuals with their high level. BLM resilience diminished, whereas in experiments with LPVLD of persons with a normal LP spectrum, BLM resistance was found to be increased. It is thus shown that despite the absence of protein receptors in the BLM some of their physico-chemical characteristics change on interaction with blood plasma LP.

The light-induced free radical ESR signals in chromatophores and reaction center preparation of R. rubrum were studied. It has been shown that the properties of the signals in both systems were strictly identical. At the microwave powers higher than 10 mW the ESR signal at g approximatley 2.00 is saturated and its asymmetry arises. Changes of the ESR signal shape may be explained by the superposition of P870+ signal with that from the ubisemiquinone. Addition of ferricyanide to chromatophores induces in the dark a new ESR signal characterized by a linewidth of 4.5 Oe which presumably arises from the antenna bacteriochlorophyll aggregates, because this signal is absent in the reaction center preparations.

Succinate synthesis from exogenous malate, alpha-ketoglutarate, oxaloacetate and L-glutamate in isolated oxygen-deprived rat heart mitochondria was studied using 1H NMR. The highest rate of succinate synthesis was observed during incubation of mitochondria with a mixture of L-glutamate and oxaloacetate. When mitochondria were incubated with [U-13C] glutamate and oxaloacetate the [U-13C] succinate/succinate and aspartate/succinate ratios were equal to 2. This suggests that the succinate produced from [U-13C] alpha-keto-glutarate formed via transamination of [U-13C] glutamate with oxaloacetate by aspartate aminotransferase exceeds twofold that synthesized via oxaloacetate reduction. It may thus be expected that GTP yield in a reaction catalyzed by the succinic thiokinase will be 2 times higher that of ATP production coupled with NADH-dependent fumarate reduction.

A decay of fluorescence probe 1-anilino-8-naphtalene sulfonate (ANS) sorbed on low density lipoproteins (LDL) surface obtained from human plasma was described. It was demonstrated that on the LDL surface the ANS probe is allocated among two pools of molecules with the time of fluorescence decay 4-7 ns and 12-16 ns. One may conclude that 75-90% of the probe is connected with lipid LDL matrix.

Tiron (1,2-dihydroxybenzene-3,5-disulfonic acid, disodium salt) was used as a spin trap to detect superoxide radicals produced by rat heart mitochondria. It was shown that ischemia results in the enhancement of the mitochondrial superoxide-forming activity. In the presence of the oxidative phosphorylation uncoupler mesoxalonitrile (3-chlorophenyl)-hydrazone the superoxide production rate in the control mitochondria increases, that in the ischemic mitochondria remains unchanged.

A fast and simple method of 1H-NMR spectroscopic control of liposome membrane integrity is suggested. The method is based on the redistribution of intensities between two singlet 1H-NMR signals--from intraliposomal marker compound (nitrilotriacetic acid sodium salt, 1H-NMR signal at 4 p.p.m.) and from its complex with Eu3+ added to the external medium (NMR signal of the complex at - 1 p.p.m.). The method permits registration of the kinetics of liposome destruction under the action of detergent or serum. It is shown that the presence of cholesterol in the membrane makes it more stable.

The rate of the antimycin-induced H2O2 and O(2) generation in beef heart submitochondrial particles is maximal at the [succinate]/[fumarate] ratio of approximately 1:5 and decays at both higher and lower redox potentials. Succinate dehydrogenase inhibitors, such as TTFA or malonate, stimulate active oxygen production in the presence of excess succinate but are inhibitory at Eh values more positive than the optimal. The modulation of O(2) and H2O2 generation by these inhibitors can be explained by their effects on the steady-state redox potential(s) of the component(s) of mitochondrial site 2.

It was shown that a short-time exposure of the spores of Act. roseolus 981 to air (oxygen) on their lyophilization by the chamber method resulted in formation of increased numbers of the free radicals in the lyophilized spores suspension and decreased the survival of the spores on their further storage under vacuum. The phenomena were less pronounced when the exposure of the spores to the air was excluded by using the collector method of lyophilization. This testified to the advantages of the latter method as compared to the chamber one. No effect of the lyophilization methods on the population composition of the spontaneous variants of Act. roseolus 981 was observed.

Apolipoprotein A-I can spontaneously associate with phosphatidylcholine and cholesterol in 2-chloroethanol-water mixture. It was demonstrated, using a spin label technique, that dissolved molecules participate in complex formation. The apolipoprotein A-I/phosphatidylcholine/cholesterol complexes were isolated by gel chromatography. Complexes of three types were prepared and characterized: type A, large heterogeneous aggregates with molecular weight 600 000, sedimentation coefficient 10 S and the following molar composition - protein/phosphatidylcholine/cholesterol, 1:(70-100):(10-12); types B and C, with weight average molecular weights 140 000 and 110 000, average sedimentation coefficients 3.6 S and 1.7 S, respectively. Both types have the same molar composition - protein/phosphatidylcholine/cholesterol, 1:25:8. The dissimilar sedimentation coefficients between complexes B and C may be explained by the difference in the monomer/tetramer ratio (monomer molecular weight 50 000). The spin label sn-1-O-stearoyl-2-O-9'-spiro(4'',4''-dimethyloxazolidine-3''-oxyl) heptadecanoylglycero-3-phosphocholine introduced into the complexes A and B showed different thermal properties of these complexes, which may be due to differences in the lipid-protein interactions.

The effect of depletion of energy stores of rat hearts on their resistance to a total of 25 min ischemia was investigated by using a 31P-NMR method. Three experimental groups were compared: (1) pyruvate-perfused hearts depleted of adenine nucleotides (35% of normal) by 2-deoxyglucose (DG) treatment and containing deoxyglucose-6-phosphate (c. 40 mumol/g dry wt); (2) hearts partially depleted of glycogen stores (40 to 50% of initial) by long-term (2h) perfusion with pyruvate; (3) glucose perfused (11 nM) hearts with normal ATP and glycogen contents. By the end of ischemia the intracellular pH was decreased by 0.33, 0.90 and 1.40 units, respectively. Time to peak of ischemic contracture increased in this series from 3 to 18 and 24 min, respectively. At the peak of ischemic contracture ATP content was c. 30 to 40% (6 to 8 mumol/g dry wt) of normal value in all three groups. Reperfusion of hearts resulted in development of significant reperfusion contracture in glucose-perfused hearts and minor contracture in other series. Recovery of high energy phosphates and cardiac work index in DG-treated, glycogen-depleted and glucose-perfused hearts were: for phosphocreatine (PCr), 72, 102 and 83%; for ATP, 29, 47 and 56% and for cardiac work, 66, 78 and 24%, respectively. Recovery of cardiac work did not correlate linearly with tissue ATP. These data demonstrate that post-ischemic recovery of the contractile function of isovolumic heart may be dissociated from pre-ischemic myocardial ATP and glycogen contents. This dissociation can be explained by the two major factors: (1) the contribution of ischemic acidosis and catabolites accumulation to the cell damage and (2) by ATP compartmentation.

The process of lyophylisation carried out very carefully under strictly defined conditions does not in inself bring about the changes in the parameters of the ESR signal with g approximately equal to 2.00 observed in animal cells. The free radicals responsible for the ESR signal observed under usual experimental conditions, result from a contact of the lyophylised specimen with moisture at an optimum water content of about 10%.

A sharp fall of ESR signal I is observed at temperature decrease while switching on light of 600 nm on the background of light of 707 nm; kinetics of the initiation of the signal under the illumination of light of 707 nm gets more complicated; the induction phase induced by previous illumination with the light of 600 nm increases.

The effect of temperature on ESR spectra of spin-labelled myosin and myosin-ADP complex was studied. It was shown that mobility of the sptrongly immobilized spin label almost did not change with temperature (-2 degrees C-37 degrees C) whereas mobility of the slightly immobilized spin label changed in a complicated manner and not monotonously. The observed changes in mobility of the slightly immobilized spin label and characteristics of the ESP spectrum of the myosin-ADP complex suggest that temperature causes continuous changes of the structure in the local environment of the binding sites of the spin label, and on the average, one conformation of myosin and myosin-ADP complex molecules exists at any temperature.

Addition of 2-deoxyglucose (DG, 8 to 13 mM) to a perfusate containing 5 mM pyruvate as oxidizable substrate caused gradual decline of contractile function of Langendorff-perfused isovolumic rat heart. Simultaneously 31P-NMR spectra showed accumulation of 2-deoxyglucose-6-phosphate (DG-6P) and decrease in phosphocreatine (PCr) and ATP contents; inosine appeared in high concentration in perfusate leaving heart. Subsequent reperfusion of the heart with DG-free solution resulted in the recovery of contractile function and PCr (to 75%), as well as in slow decay of DG-6P at unchanged low level of ATP (35%). No correlation was found between tissue ATP content and contractile function, when expressed as the product of developed pressure and heart rate. In contrast, contraction correlated with tissue PCr level at low ATP. These data demonstrate that effective contraction of the isovolumic heart is possible at substantially decreased level of cytoplasmic ATP. The results of this study are in accord with a concept of ATP compartmentation in cardiac cells.

It is shown that three types of ESR spectra of myosin labeled with both acetate and acetamide spin labels are distinguished. These three types are the spectrum of initial preparation, myosin--ADP complex spectrum and the spectrum recorded during ATP hydrolysis. The ESR spectra of myosin labeled with acetate spin label are more sensitive to conformational changes of myosin induced by complex formation and temperature.

Inhibition of electron transport in bovine heart submitochondrial particles resulting from treatment with British antilewisite is accompanied by suppression of superoxide radical and H2O2 generation at the b-c1 site of the respiratory chain. In conjunction with the previously obtained data, the results suggest that the effect of British antilewisite is directed towards the step of ubihydroqinone oxidation to ubisemiquinone by the Rieske FeS-protein.

The effects of two molecular forms of water-soluble ferredoxin (Fd I and Fd II) on the kinetics of electron transport in bean chloroplasts (class B) were studied. The light-induced redox transitions of the photosystem I reaction center P700 were measured by the intensity of the EPR signal I produced by P700+. Both forms of ferredoxin, Fd I and Fd II, when added to the chloroplasts in catalytic amounts, stimulate the light-induced electron transfer from P700 to NADP+. Nevertheless, Fd I is a better mediator of the back reactions from NADPH to P700+. This electron transfer pathway is sensitive to the cyclic electron transport inhibitor, antimycin A, and to DCMU inhibitor of electron transport between photosystem II and plastoquinone. It may be concluded that the two molecular forms of ferredoxin, Fd I and Fd II, differ in their ability to catalyze cyclic electron transport in photosystem I. The role of Fd I and Fd II in regulation of electron transport at the acceptor site of photosystem I is discussed.

The effect of ischemia on the formation of products of anaerobic metabolism and their release into the cardiac effluent in isolated perfused guinea pig hearts was studied. During 30 min normothermal ischemia, the myocardial ATP and phosphocreatine levels decreased to 34% and 15% of the initial values, respectively. The net alanine formation in ischemia was approximately a stoichiometric glutamate decrease; the increase in the tissue malate content corresponded to the aspartate----oxaloacetate----malate anaplerotic flux, the succinate production being commensurable to alpha-ketoglutaric acid formation in the alanine aminotransferase reaction. Using 1H-NMR, it was shown that the release of trace amounts of lactate, alanine, succinate, creatine and pyruvate into cardiac effluents occurred during the first 5 minutes of reperfusion. The rate of metabolite release decreased in the following order: lactate much greater than alanine greater than succinate greater than creatine. By the 30th minute of reperfusion, the decrease in the tissue levels of these metabolites to preischemic values was accompanied by the recovery of ATP and phosphocreatine to 65% and 90% of the initial levels, respectively. The data obtained suggest that the formation and release of alanine, creatine or succinate as well as lactate from ischemic myocardium may testify to significant disturbances in energy metabolism of the myocardium.

Thermo-induced structural reorganization in human high density lipoproteins subclasses HDL2 and HDL3 have been studied by means of electronic paramagnetic resonance method. A number of spin labels were used: 5- and 16-DOXYL stearates, DOXYL-labelled derivatives of pentadecane, phosphatidyl choline and sphingomyelin. In hypoalphalipoproteinemia one of the thermo-induced transitions in HDL2 (at 23-30 degrees) was not observed. In coronary heart disease the critical temperature for HDL3 was shifted from the physiological values and located at 42-48 degrees. The data obtained are discussed from the point of view of their potential role in atherogenesis.

Fluorescent probe I-aniline-8-naphthalene sulfonate was used to evaluate the LDL surface potential values in patients with coronary artery atherosclerosis. The more negative charge was found on the LDL surface in patients with atherosclerosis of three coronary arteries as compared with that of healthy persons.

Tiron (1,2-dihydroxybenzene-3,5-disulfonate) is oxidized to an EPR-visible semiquinone by superoxide radicals produced by xanthine oxidase. The steady-state level of the Tiron radicals increases with an increased xanthine oxidase concentration. A calibration plot has been obtained relating the steady-state concentration of the Tiron semiquinone determined by EPR-spectroscopy to the rate of 0.2 production as measured by the superoxide dismutase-sensitive cytochrome c reduction. This approach allows for a simple and sensitive assay of 0.2 generation rate in biological systems in the range of ca.0.1-4.0 microM/min using Tiron as a spin trap. The rate of 0.2 generation by antimycin-inhibited ischemic rat heart mitochondria has been measured by this method.

Using 31P-NMR technique it was shown that exogenous phosphocreatine (10 mM) added in the cardioplegic solution provided higher levels of recovery of intracellular ATP (approx. 60% vs. 26% in the control) and phosphocreatine (90% vs. 43%) in perfused rat hearts after 35 min of total ischemia. Simultaneously significantly higher levels of contractile recovery (90% vs. 35%) and a three-fold decrease in creatine kinase release into the perfusate were observed. These effects of exogenous phosphocreatine can be related to either intracellular action of this compound or its interaction with cellular membrane.

The method of atherogenic class lipoproteins fractionation by equilibrium zonal centrifugation in SW 41 Ti rotor (density range 1,006-1,063 g/ml) was proposed. Hydrated densities, flotation coefficients and mean diameters of LDL particles from different subfractions were estimated.

Sodium ion interaction with sarcoplasmic reticulum (SR) membranes leads to considerable alterations of the [23Na]NMR lineshape. Na+ binding to SR in the presence of Ca2+ and H+ is well described by a model which postulates a competitive ion binding to high and low affinity sites of Ca2+-ATPase. The dissociation constant, Kd, for high and low affinity sites is 5 and 10 mM, respectively, for Na+ and (3-5).10(-8) and 1.5.10(-3) M, respectively, for Ca2+. The pK value for high and low affinity sites is 7.3 and 6.1, respectively. Other alkaline metal ions compete with Na+ for the low affinity sites of Ca2+-ATPase; their affinities decrease in the following order: Na+ = K+ greater than Rb+ greater than Cs greater than Li+. Some of the Na+ binding sites (approximately 10%) do not interact with Ca2+.

The lipid membrane structure of two myxoviruses (influenza and Sendai viruses), red blood cells and liposomes (prepared from total lipids of Sendai virus) have been investigated in the presence of rimantadine by means of two spin probes C5 and C6--the iminoxyl derivatives of stearic acids. Rimantadine was shown to penetrate the lipid membranes of myxoviruses and red blood sells and change the structure of the lipid phase no less than 0.8 nm apart from the viral surface. In the depth of 2.2 nm the structure of these lipid membranes remains unchanged after addition of rimantadine. Rimantadine also displaces the phase transition points of cell and viral lipid membranes to lower temperatures. Rimantadine does not change the lipid bilayer structure of liposomes 0.8 nm apart from its surface. These finding allow to suggest that rimantadine affects the cell and viral lipoprotein membranes.

After the adaptation of leaves to darkness (2--4 min) the kinetics of the initiation of ESR I signal aquires a multiphase character: after an initial leap the stage of slower growth sets in. The "memory" of the system about the effect of light of 600 nm, which was realized as a delay of the growth of the ESR I signal under the effect of light of 707 nm, was preserved in the darkness for several minutes. If the light of 600 nm is switched on at the background of the light of 707 nm not earlier than 2-4 min after its switching off in the preceding cycle, unmonotonous transitional process with an overleap is observed.

It has been found in experiments with high resolution 31P-NMR spectroscopy (200 MHz) that the phosphocreatine peak is splitted into two different peaks in the mixtures of H2O and D2O and is single but with different chemical shifts in pure H2O and D2O. This phenomenon is explained by substitution of protons of guanidino group in phosphocreatine by deuterium. The effect of splitting disappeared at extreme pH values (greater than 8.5 or less than 4.0) and at temperatures higher than 45 degrees C due to accelerated proton-deuterium exchange. Creatine kinase added to phosphocreatine solution also lowered its temperature of peaks' collapse by 5 degrees-10 degrees C. A saturation (spin) transfer method was used to show that the phosphoryl group transfer to ADP in creatine kinase active center is slower with deuterium-substituted phosphocreatine than with H-phosphocreatine. The data are taken to show the importance of the proton transfer step in the creatine kinase reaction mechanism and acceleration of phosphocreatine proton-deuterium exchange by creatine kinase.

Electron spin resonance (ESR) spectra of the spin probes C5 and C16, the iminoxyl derivatives of stearic acids, have been investigated after their incorporation into lipid membranes of influenza and Sendai viruses. At room temperature the spectra of both probes coincided indicating the similar composition and origin of lipid viral membranes. Similar to influenza virus, Sendai virus was shown to possess the external lipid bilayer. Investigation of the rigidity of the lipid phase of myxoviruses as a function temperature has shown that the structure of the lipid external layer (no less than 8 A in thickness) is due to surface glycoproteins incorporated into the viral membrane. The structural transition of the lipid phase of influenza and Sendai viruses 8 A apart from the viral surface was observed at 50 degrees C. The surface glycoproteins do not probably penetrate deep in the lipid membrane of myxoviruses and do not condition its structure at long distances (22 A) from the viral surface. The completely reversible structural transition of lipid phase at this distance was observed at temperatures 17--20 degrees C.