Effect of lanthanides on oxidation of Mn2+ cations via a high-affinity Mn-binding site in photosystem II membranes.

Lanthanide cations (La3+ and Tb3+) bind to the Ca-binding site of the oxygen-evolving complex in Ca-depleted PSII membranes and irreversibly inhibit the oxygen evolution. Оn the other hand, EPR measurement of Mn2+ concentration in buffer revealed that lanthanide cations inhibit the light-dependent oxidation of Mn2+ cations via the high-affinity Mn-binding site in Mn-depleted PSII membranes, which suggests that they bind to and inhibit the high-affinity Mn-binding site of the oxygen-evolving complex. The inhibition is irreversible, bound Ln3+ cation could not be washed out from the sample. Calcium ion inhibits oxidation of Mn2+ (5 µM) at very high concentration (tens mM) and the inhibition is reversible. In this work we measured the reduction rate of exogenic electron acceptor 2,6-dichlorophenolindophenol during the oxidation of Mn2+ cations in the Ca-depleted PSII and in the Ca-depleted PSII treated with lanthanides after extraction of Mn cluster from these preparations. We found that irreversible binding of the lanthanide cation to the Ca-binding site in the Ca-depleted PSII membranes leads to a partial inhibition of the high-affinity Mn-binding site.

pH dependence of photosystem II photoinhibition: relationship with structural transition of oxygen-evolving complex at the pH of thylakoid lumen.

Ca-depleted photosystem II membranes (PSII[-Ca]) do not contain PsbP and PsbQ proteins protecting the Mn4CaO5 cluster of the PSII oxygen-evolving complex (OEC). Therefore, the Mn ions in the PSII(-Ca) membranes can be reduced by exogenous bulky reductants or the charged reductant Fe(II). We have recently found that the resistance of Mn ions in the OEC to the Fe(II) action is pH dependent and that this reductant is less effective at pH 5.7 than at pH 6.5 (Semin et al. J Photochem Photobiol B 178:192, 2018). Taking these data into account, we investigated the photoinhibition in different PSII membranes at pH 5.7 and 6.5 and found that the resistance to photoinhibition of PSII and PSII(-Ca) membranes with a Mn cluster is higher at pH 5.7 than at pH 6.5, whereas the resistance of the Mn-depleted PSII membranes is pH independent. In thylakoids, light generates the transmembrane ΔpH, leading to the acidulation of lumen that results in pH 5.7. The uncouplers (NH4Cl or nigericin) that significantly prevent acidulation increase the rate of PSII photoinhibition in thylakoids. We suggest that the structural transition in the OEC at pH 5.7 plays a role of a built-in mechanism increasing the resistance of OEC to photoinhibition under illumination, since it is accompanied by a pH decrease in lumen to 5.7. The coincidence of these pH values, i.e. lumen pH under illumination and pH of the maximal resistance of the Mn cluster to the reduction by reductants, can point at the pH-dependent mechanism of PSII self-protection from photoinactivation.

Mechanism of inhibition and decoupling of oxygen evolution from electron transfer in photosystem II by fluoride, ammonia and acetate.

Ca(2+) extraction from oxygen-evolving complex (OEC) of photosystem II (PSII) is accompanied by decoupling of oxygen evolution/electron transfer processes [Semin et al. Photosynth. Res. 98 (2008) 235] and appearance of a broad EPR signal at g=2 (split "S3" signal) what can imply the relationship between these effects. Split signal have been observed not only in Ca-depleted PSII but also in PSII membranes treated by fluoride anions, sodium acetate, and NH4Cl. Here we investigated the question: can such compounds induce the decoupling effect during treatment of PSII like Ca(2+) extraction does? We found that F(-), sodium acetate, and NH4Cl inhibit O2 evolution in PSII membranes more effectively than the reduction of artificial electron acceptor 2,6-dichlorophenolindophenol, i.e. the action of these compounds is accompanied by decoupling of these processes in OEC. Similarity of effects observed after Ca(2+) extraction and F(-), CH3COO(-) or NH4Cl treatments suggests that these compounds can inactivate function of Ca(2+). Such inactivation could originate from disturbance of the network of functionally active hydrogen bonds around OEC formed with participation of Ca(2+). This inhibition effect is observed in the region of low concentration of inhibitors. Increasing of inhibitor concentration is accompanied by appearance of other sites of inhibition.

Rapid degradation of the tetrameric Mn cluster in illuminated, PsbO-depleted photosystem II preparations.

A "decoupling effect" (light-induced electron transport without O2 evolution) was observed in Ca-depleted photosystem II (PSII(-Ca)) membranes, which lack PsbP and PsbQ (Semin et al. (2008) Photosynth. Res., 98, 235-249). Here PsbO-depleted PSII (PSII(-PsbO)) membranes (which also lack PsbP and PsbQ) were used to examine effects of PsbO on the decoupling. PSII(-PsbO) membranes do not reduce the acceptor 2,6-dichlorophenolindophenol (DCIP), in contrast to PSII(-Ca) membranes. To understand why DCIP reduction is lost, we studied light effects on the Mn content of PSII(-PsbO) samples and found that when they are first illuminated, Mn cations are rapidly released from the Mn cluster. Addition of an electron acceptor to PSII(-PsbO) samples accelerates the process. No effect of light was found on the Mn cluster in PSII(-Ca) membranes. Our results demonstrate that: (a) the oxidant, which directly oxidizes an as yet undefined substrate in PSII(-Ca) membranes, is the Mn cluster (not the Y(Z) radical or P680(+)); (b) light causes rapid release of Mn cations from the Mn cluster in PSII(-PsbO) membranes, and the mechanism is discussed; and (c) rapid degradation of the Mn cluster under illumination is significant for understanding the lack of functional activity in some PSII(-PsbO) samples reported by others.

Investigation of the low-affinity oxidation site for exogenous electron donors in the Mn-depleted photosystem II complexes.

In the manganese-depleted photosystem II (PSII[-Mn]) preparations, oxidation of exogenous electron donors is carried out through the high-affinity (HA) and the low-affinity (LA) sites. This paper investigates the LA oxidation site in the PSII(-Mn) preparations where the HA, Mn-binding site was blocked with ferric cations [[11] B.K. Semin, M.L. Ghirardi, M. Seibert, Blocking of electron donation by Mn(II) to Y(Z)(*) following incubation of Mn-depleted photosystem II membranes with Fe(II) in the light, Biochemistry 41 (2002) 5854-5864.]. In blocked (PSII[-Mn,+Fe]) preparations electron donation by Mn(II) cations to Y(Z)(*) was not detected at Mn(II) concentration 10 microM (corresponds to K(m) for Mn(II) oxidation at the HA site), but detected at Mn concentration 100 microM (corresponds to K(m) for the LA site) by fluorescence measurements. Comparison of pH-dependencies of electron donation by Mn(II) through the HA and the LA sites revealed the similar pK(a) equal to 6.8. Comparison of K(m) for diphenylcarbazide (DPC) oxidation at the LA site and K(d) for A(T) thermoluminescence band suppression by DPC in PSII(-Mn,+Fe) samples suggests that there is relationship between the LA site and A(T) band formation. The role of D1-His190 as an oxidant of exogenous electron donors at the LA site is discussed. In contrast to electrogenic electron transfer from Mn(II) at the HA site to Y(Z)(*), photovoltage due to Mn(II) oxidation in iron-blocked PSII(-Mn) core particles was not detected.

Effect of calcium chelators on the formation and oxidation of the slowly relaxing reduced plastoquinone pool in calcium-depleted PSII membranes. Investigation of the F0 yield.

The F(0) fluorescence yield in intact photosystem II (PSII), Ca-depleted PSII (PSII(-Ca/NaCl)), and Mn-depleted PSII membranes was measured before and after dim light treatment (1-2 min), using flash-probe fluorescence and fluorescence induction kinetic measurements. The value of F(0) after the light treatment (F'(0)) was larger than F(0) in dark-adapted PSII membranes and depended on the appearance of the slowly relaxing, reduced plastoquinone pool (t(1/2) = 4 min) formed during preillumination, which was not totally reoxidized before the F'(0) measurement. In PSII(-Ca/NaCl) such a pool also appeared, but the F'(0) yield was even higher than in intact PSII membranes. In Mn-depleted PSII membranes, the pool did not form. Interestingly, the yield of F'(0) in Ca-depleted PSII membranes prepared using chelators (EGTA and citrate) or containing 5 mM EGTA was significantly lower than in PSII(-Ca/NaCl) samples prepared without chelators. These data indicate that chelators inhibit the reduction of Q(A) and Q(B) and formation of the slowly relaxing plastoquinone pool, or alternatively they increase the rate of its oxidation. Such an effect can be explained by coordination of the chelator molecule to the Mn cluster in PSII(-Ca/NaCl) membranes, rather than different amounts of residual Ca2+ in the membranes (with or without the chelator), since the remaining oxygen-evolving activity (approximately 15%) in PSII(-Ca/NaCl) samples did not depend on the presence of the chelator. Thus, chelators of calcium cations not only have an effect on the EPR properties of the S2 state in PSII(-Ca/NaCl) samples, but can also influence the PSII properties determining the rate of plastoquinone pool reduction and/or oxidation. The effect of some toxic metal cations (Cd, Cu, Hg) on the formation of the slowly relaxing pool in PSII membranes was also studied.

pH dependence of the efficiency of binding of iron cations to the donor side of photosystem II.

Light-induced interaction of Fe(II) cations with the donor side of Mn-depleted photosystem II (PS II(-Mn)) results in the binding of iron cations and blocking of the high-affinity (HAZ) Mn-binding site. The pH dependence of the blocking was measured using the diphenylcarbazide/2,6-dichlorophenolindophenol test. The curve of the pH dependence is bell-shaped with pK1 = 5.8 and pK2 = 8.0. The pH dependence of the O2-evolution mediated by PS II membranes is also bell-shaped (pK2 = 7.6). The pH dependence of the process of electron donation from exogenous donors in PS II(-Mn) was studied to determine the location of the alkaline pH sensitive site of the electron transport chain. The data of the study showed that the decrease in the iron cation binding efficiency at pH > 7.0 during blocking was determined by the donor side of the PS II(-Mn). Mössbauer spectroscopy revealed that incubation of PS II(-Mn) membranes in a buffer solution containing 57Fe(II) + 57Fe(III) was accompanied by binding only Fe(III) cations. The pH dependence of the nonspecific Fe(III) cation binding is also described by the same bell-shaped curve with pK2 = 8.1. The treatment of the PS II(-Mn) membranes with the histidine modifier diethylpyrocarbonate resulted in an increase in the iron binding strength at alkaline pH. It is suggested that blocking efficiency at alkaline pH is determined by competition between OH- and histidine ligand for Fe(III). Because the high-affinity Mn-binding site contains no histidine residue, this fact can be regarded as evidence that histidine is located at another (other than high-affinity) Fe(III) binding site. In other words, this means that the blockage of the high-affinity Mn-binding site is determined by at least two iron cations. We assume that inactivation of oxygen-evolving complex and inhibition of photoactivation in the alkaline pH region are also determined by competition between OH- and a histidine residue involved in coordination of manganese cation outside the high-affinity site.

Comparative study of effects of artificial electron donors on the AT-band of photosystem II thermoluminescence.

Extraction of the Mn-cluster from photosystem II (PS II) inhibits the main bands of thermoluminescence and induces a new AT-band at -20 degrees C. This band is attributed to the charge recombination between acceptor QA- and a redox-active histidine residue on the donor side of PS II. The effect of Mn(II) and Fe(II) cations as well as the artificial donors diphenylcarbazide and hydroxylamine on the AT-band of thermoluminescence was studied to elucidate the role of the redox-active His residue in binding to the Mn(II) and Fe(II). At the Mn/PS II reaction center (RC) ratio of 90 : 1 and Fe/PS II RC ratio of 120 : 1, treatment with Mn(II) and Fe(II) causes only 60% inhibition of the AT-band. Preliminary exposure of Mn-depleted PS II preparations to light in the presence of Mn(II) and Fe(II) causes binding of the cations to the high-affinity Mn-binding site, thereby inhibiting oxidation of the His residue involved in the AT-band formation. The efficiency of the AT-band quenching induced by diphenylcarbazide and hydroxylamine is almost an order of magnitude higher than the quenching efficiency of Mn(II) and Fe(II). Our results suggest that the redox-active His is not a ligand of the high-affinity site and does not participate in the electron transport from Mn(II) and Fe(II) to YZ. The concentration dependences of the AT-band inhibition by Mn(II) and Fe(II) coincide with each other, thereby implying specific interaction of Fe(II) with the donor side of PS II.

Photosystem II of peas: effects of added divalent cations of Mn, Fe, Mg, and Ca on two kinetic components of P(+)(680) reduction in Mn-depleted core particles.

The catalytic Mn cluster of the photosynthetic oxygen-evolving system is oxidized via a tyrosine, Y(Z), by a photooxidized chlorophyll a moiety, P(+)(680). The rapid reduction of P(+)(680) by Y(Z) in nanoseconds requires the intactness of an acid/base cluster around Y(Z) with an apparent functional pK of <5. The removal of Mn (together with bound Ca) shifts the pK of the acid/base cluster from the acid into the neutral pH range. At alkaline pH the electron transfer (ET) from Y(Z) to P(+)(680) is still rapid (<1 micros), whereas at acid pH the ET is much slower (10-100 micros) and steered by proton release. In the intermediate pH domain one observes a mix of these kinetic components (see R. Ahlbrink, M. Haumann, D. Cherepanov, O. Bögershausen, A. Mulkidjanian, W. Junge, Biochemistry 37 (1998)). The overall kinetics of P(680)(+) reduction by Y(Z) in Mn-depleted photosystem II (PS II) has been previously shown to be slowed down by divalent cations (added at >10 microM), namely: Mn(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+) (C.W. Hoganson, P.A. Casey, O. Hansson, Biochim. Biophys. Acta 1057 (1991)). Using Mn-depleted PS II core particles from pea as starting material, we re-investigated this phenomenon at nanosecond resolution, aiming at the effect of divalent cations on the particular kinetic components of P(+)(680) reduction. To our surprise we found only the slower, proton steered component retarded by some added cations (namely Co(2+)/Zn(2+)>Fe(2+)>Mn(2+)). Neither the fast component nor the apparent pK of the acid/base cluster around Y(Z) was affected. Apparently, the divalent cations acted (electrostatically) on the proton release channel that connects the oxygen-evolving complex with the bulk water, but not on the ET between Y(Z) and P(+)(680), proper. Contrastingly, Ca(2+) and Mg(2+), when added at >5 mM, accelerated the slow component of P(+)(680) reduction by Y(Z) and shifted the apparent pK of Y(Z) from 7.4 to 6.6 and 6.7, respectively. It was evident that the binding site(s) for added Ca(2+) and Mg(2+) were close to Y(Z) proper. The data obtained are discussed in relation to the nature of the metal-binding sites in photosystem II.

[Mossbauer spectroscopy of cells of cyanobacteria Synechocystis sp. PCC 6803 devoid of photosystem I and containing inactive phycobilisomes]. / Messbauérovskaia spektroskopiia kletok tsianobakterii Synechocystis sp. PCC 6803 bez fotosistemy I i s neaktivnymi fikobilisomami.

Mossbauer spectra of the psaAB mutant of Synechocystis sp. PPC 6803 devoid of photosystem I grown in a 57Fe-containing medium were measured. The spectrum is a broadened doublet whose size (about 20%) and parameters (isomeric shift delta = 0.3 mm/s and quadrupole splitting delta = 0.8 mm/s) suggest the presence of abundant nanoclusters of Fe3+ oxides in a superparamagnetic state tightly bound to the membrane. Treatment of cells with EDTA was accompanied by a substantial (tenfold) decrease in the amount of iron nonspecifically bound to the membrane and the appearance of Fe2+ localized, probably, inside cells and/or cell membranes. In addition, the spectrum of washed cells exhibited superfine magnetic splitting due to iron oxide clusters greater in size than nanoclusters present in the membrane prior to EDTA treatment.

Comparative study of thermal degradation of iron-sulfur proteins in spinach chloroplasts and membranes of thermophilic cyanobacteria: mössbauer spectroscopy.

Mössbauer spectra of chloroplasts isolated from spinach plants grown in a mineral medium enriched with 57Fe and Mössbauer spectra of native membranes of the thermophilic cyanobacterium Synechococcus elongatus contain a broad asymmetric doublet typical of the iron-sulfur proteins of Photosystem (PS) I. Exposure of chloroplasts to temperatures of 20-70 degrees C significantly modifies the central part of the spectra. This spectral change is evidence of decreased magnitude of the quadrupole splitting. However, the thermally induced doublet (DeltaQ = 3.10 mm/sec and delta = 1.28 mm/sec) typical of hydrated forms of reduced (divalent) inorganic iron is not observed in spinach chloroplasts. This doublet is usually associated with degradation of active centers of ferredoxin, a surface-exposed protein of PS I. The Mössbauer spectra of photosynthetic membranes of spinach chloroplasts and cyanobacteria were compared using the probability distribution function of quadrupole shift (1/2 quadrupole splitting DeltaQ) of trivalent iron. The results of calculation of these functions for the two preparations showed that upon increasing the heating temperature there was a decrease in the probability of the presence of native iron-sulfur centers FX, FA, and FB (quadrupole shift range, 0.43-0.67 mm/sec) in heated preparations. This process was also accompanied by an increase in the probability of appearance of clusters of trivalent iron. This increase was found to be either gradual and continuous or abrupt and discrete in photosynthetic membranes of cyanobacteria or spinach chloroplasts, respectively. The probability of the presence of the iron-sulfur centers FX, FA, and FB in chloroplasts abruptly decreases to virtually to zero within the temperature range critical for inhibition of electron transport through PS I to oxygen. In cyanobacteria, both thermal destruction of iron-sulfur centers of PS I and functional degradation of PS I are shifted toward a higher temperature. The results of this study suggest that the same mechanism of thermal destruction of the PS I core occurs in both thermophilic and mesophilic organisms: destruction of iron-sulfur centers FX, FA, and FB, release of oxidized (trivalent) iron, and its accumulation in membrane-bound iron-oxo clusters.

Inorganic Fe2+ formation upon Fe-S protein thermodestruction in the membranes of thermophilic cyanobacteria: Mössbauer spectroscopy study.

A model description of the Mössbauer spectrum (80 K) of native membranes of the thermophilic cyanobacterium Synechococcus elongatus is suggested on the basis of the known values of quadrupole splitting (deltaE(Q)) and isomer shift (deltaFe) for the iron-containing components of the photosynthetic apparatus. Using this approach, we found that heating the membranes at 70-80 K results in a decrease of doublet amplitudes belonging to F(X), F(A), F(B) and ferredoxin and simultaneous formation of a new doublet with deltaE(Q) = 3.10 mm/s and delta-Fe = 1.28 mm/s, typical of inorganic hydrated forms of Fe2+. The inhibition of electron transfer via photosystem I to oxygen, catalyzed by ferredoxin, occurs within the same range of temperatures. The data demonstrate that the processes of thermoinduced Fe2+ formation and distortions in the photosystem I electron transport in the membranes are interrelated and caused mainly by the degradation of ferredoxin. The possible role of Fe2+ formation in the damage of the photosynthetic apparatus resulting from heating and the action of other extreme factors is discussed.

Coordination sphere and structure of the Mn cluster of the oxygen-evolving complex in photosynthetic organisms.

The great similarity between the binding of Fe(II) and the high-affinity Mn-binding site in the Mn-depleted PSII membranes (Semin et al. (1996) FEBS Lett. 375, 223-226) suggests that the coordination sphere of Mn in PSII is also suitable for iron. A comparison is performed between the primary amino acid sequences of D1 and D2 and diiron-oxo enzymes with the function of oxygen activation. All conservative motifs (EXXH) and residues binding and stabilizing the diiron cluster in diiron-oxo enzymes have been found in the C-terminal domains of D1 and D2 polypeptides. On the basis of these sequence similarities we suggest a structural model for the manganese cluster in the oxygen-evolving complex.

High-specific binding of Fe(II) at the Mn-binding site in Mn-depleted PSII membranes from spinach.

The interaction of Fe(II) and Fe(III) with the 'high-affinity Mn-binding site' in Mn-depleted photosystem II (PSII) was investigated using diphenilcarbazide (DPC)/2,6-dichlorophenol-indophenol (DCIP) inhibition assay. Fe(III) was ineffective in the inhibition of DPC-DCIP reaction while Fe(II) decreased the rate of DCIP photoreduction supported by DPC in the same concentration range as Mn(II). The effectivity of the interaction of Fe(II) with the high affinity Mn-binding site depends on different anions in the same manner as for Mn(II) and coincides with hierarchy observed for the stimulation of O2 evolution. The Fe(II) binding is accompanied by its oxidation. By using reductants it was shown that the high affinity site contains a redox-active component and the reduction of this component totally prevents the binding of Fe(II).

Effect of formate on Mössbauer parameters of the non-heme iron of PS II particles of cyanobacteria.

Mössbauer spectra were measured for PSII particles having an active water-splitting system. The particles were isolated from the thermophilic cyanobacterium Synechococcus elongatus enriched in 57Fe. The Mössbauer resonance absorption spectrum is a superposition of 3 doublets with the following quadrupole splitting and chemical shift: 1, delta = 0.40, delta = 0.85; II, delta = 1.35, delta = 2.35; III, delta = 0.25, delta = 1.65. The delta and delta values of doublets I, II, III are characteristic of proteins with iron-sulphur center, non-heme iron of the reaction center of higher plants and of the oxidized cytochrome b-559. Treatment with sodium formate to remove bicarbonate affects only the doublet of non-heme iron, causing its quadrupole splitting to reduce to 1.75 and the chemical shift to reduce to 0.90. After washing out the formate, the Mössbauer spectrum of non-heme iron is restored. The data suggest that bicarbonate is a ligand for the non-heme iron of the reaction center of cyanobacteria.

Tetracaine inhibition of electron transport in pea chloroplasts is coupled to calcium displacement by the local anesthetic.

Chloroplasts isolated from pea seadlings grown on water containing 45Ca2+ were treated with local anesthetic tetracaine. Addition of tetracaine inactivated the electron transport activity of donor side photosystem II. This inhibition was accompanied by 45Ca2+ release from the chloroplast membranes as the whole and destroyed by osmotic shock. No such effect was observed when Tris or hydroxylamine were used to inhibit the donor side photosystem II. Upon thermal inactivation of chloroplasts 45Ca2+ release occurred but at temperatures above 80 degrees. The functional role of Ca2+ in photosystem II is discussed.

[Comparative analysis of sequences of protein D2 from photosystem 2 reaction center and various Ca 2+-binding proteins]. / Sravnitel'nyi analiz posledovatel'nostei belka D2 reaktsionnogo tsentra fotosistemy 2 i nekotorykh Ca 2+-sviazyvaiushchikh belkov.

The amino acid sequence of D2 protein was compared with those of calcium binding proteins and receptor for calcium channel blockers in connection with the data showing the participation of Ca2+ in photosystem 2 electron transport and the inhibition of this process by calmodulin antagonists, calcium channel blockers and local anesthetics. Protein D2 possesses a pattern analogous to the "EF-hand" sites of the calcium binding proteins. Comparison of the amino acid sequence of the calmodulin fragment binding the phenothiazine type calmodulin antagonists with the amino acid sequence of D2 protein and calcium channel protein revealed a high degree of sequence identity. Common structural features take place also between the membrane spanning segment III of D2 protein, which contains the tyrosine residue (161), responsible for ESR-signal IIS, and the membrane segment IVS5 of calcium channel protein. A model explaining the mechanism of calcium function in the oxygen-evolving system is proposed.

Local anesthetics-induced inhibition of chloroplast electron transport.

The effects of local anesthetics on photosynthetic activity of pea chloroplasts were investigated in order to elucidate the role of Ca2+ in photosynthetic electron transport. Dibucaine, benzocaine and tetracaine were found to inhibit the O2-evolving activity. The inhibitory effect decreases in the order dibucaine greater than benzocaine greater than tetracaine greater than trimecaine similarly as does the potency to inhibit propagation of excitation in nerve fibre. As demonstrated in experiments with artificial donors and acceptors, the site of inhibition is the water-splitting site of PSII. The inhibitory power of the anesthetics grows with increasing ionic strength of the incubating mixture (by adding NaCl or MgCl2) and with pH; this is explained by occurrence of the neutral form of amine. At low concentrations the charged anesthetic acts as a protonofore; however, the inactivation of water splitting is not due to the protonophoric effect. The incubation is followed by the disappearance of ESR signal IIs. The role of Ca2+ and Ca2+-binding protein in PSII electron transport and its localization are discussed.

[The action of membranotropic compounds with various structures on the parameters of the phase transition of dimyristoylphosphatidylcholine]. / Deistvie membranotropnykh soedinenii razlichnoi struktury na parametry fazovogo perekhoda dimiristoilfosfatidilkholina.

The effect of substances of different nature on the thermodynamic characteristics of dimyristoylphosphatidylcholine (DMPC) phase transition by the differential scanning microcalorimetry has been studied. The substances disposed in hydrophobic part of membrane--alpha-tocopherol, ubiquinone Q10, ionol and vitamin K3 cause the decrease of enthalpy and cooperativity of phase transition. The substances which have the side hydrocarbon chain (tocopherol and ubiquinone Q10) compared with ones without it (ionol and vitamin K3) and reduced quinones (Q10 and vitamin K3) compared with the oxidized ones have stronger influence on the enthalpy and cooperativity of transition. The inclusion of the local anesthetic dicaine disposed mainly in the zone of polar heads of phospholipids into DMPC membranes decreases the temperature of phase transition considerably and practically does not change the cooperativity. A possibility to use the method of differential scanning microcalorimetry to estimate the localization of membrane tropic substances within lipid bilayer is under discussion.

[A study of the dynamic properties of membrane proteins using Mössbauer spectroscopy]. / Issledovanie dinamicheskikh svoistv membrannykh belkov metodom messbauérovskoi spektroskopii.

While studying the parameters of "narrow" and "broad" lines appearing in Mössbauer spectra of undehydrated membrane proteins heated from 80 to 280 K it has been for the first time found for proteins that the behavior of the complete area of spectrum S does not differ from that of Debye-Waller factor. An abrupt decrease of quadrupole splitting value from delta = 0.7 mm/s to delta = 0 within the temperature range 220-270 K. Computation of the spectra with their division into 3 components responding respectively by heat, diffusion and conformational movement made possible explanation of all the evolutionary changes proceeding in them with the temperature rise. Preservation of the complete area of the spectrum S (T) is conditioned by the increase of the component responsive to conformational changes of Fe atom within 230-270 K. These movements "suppress" quadrupole splitting observed in the spectra at low temperatures. Dynamic mobility is considered in terms of the Fe atom movement in the biphase potential.