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Biomedical subjects

V D Samuilov

Publications and source records attributed to V D Samuilov.

At least 37 records · Page 2Linked to original sources

Quenching of chlorophyll fluorescence by quinones.

Quinones caused quenching of Chl a fluorescence in native and model systems. Menadione quenched twofold the fluorescence of Chl a and BChl a in pea chloroplasts, chromatophores of purple bacteria, and liposomes at concentrations of 50-80 microM. To obtain twofold quenching in Triton X-100 micelles and in ethanol, the addition of 1.3 mM and 11 mM menadione was required, respectively. A proportional decrease in the lifetime and yield of Chl a fluorescence in chloroplasts, observed as the menadione concentration increased, is indicative of the efficient excitation energy transfer from bulk Chl to menadione. The decrease in the lifetime and yield of fluorescence was close to proportional in liposomes, but not in detergent micelles. The insensitivity of the menadione quenching effect to DCMU in chloroplasts, and similarity of its action in chloroplasts and liposomes indicate that menadione in chloroplasts interacts with antenna Chl, i.e., nonphotochemical quenching of fluorescence occurs.

Bacterial Chromatophores↗

Photoreduction of silicomolybdate in chloroplasts by agents accelerating the deactivation reactions of the water-oxidizing system.

Uncouplers of photosynthetic phosphorylation, CCCP, TTFB and PCP, inhibited light-induced O2 evolution in the Hill reaction with SiMo (I50 approximately 20, 3 and 45 microM, respectively), but only insignificantly diminished SiMo photoreduction by pea chloroplasts. The same properties were exhibited by the ADRY agent ANT2p. CCCP, TTFB and PCP are oxidizable compounds with redox potentials of +1.17, +1.18 and +1.09 V (pH 6.0), as determined by cyclic voltammetry. Similarly to NH2OH, the tested uncouplers can apparently serve as electron donors for photosystem II.

Chloroplasts↗

Electrogenicity at the secondary quinone acceptor site of cyanobacterial photosystem II.

Flash-induced generation of the electric potential difference (delta psi) by a direct electrometrical method was studied in Anacystis nidulans photosystem II-containing proteoliposomes associated with a phospholipid-impregnated collodion film. Besides a rapid phase of delta psi generation corresponding to charge separation between P680 and QA, an additional electrogenic phase with a characteristic time of 0.27 ms at pH 7.0 was observed after the second laser flash. The maximal amplitude of this phase was approx. 4% of that related to the P680+QA- formation. The sensitivity of this phase to DCMU, the flash-number dependence of its amplitude as well as the amplitude and the rate constant pH-dependences, indicate that it is due to the dismutation of QA- and QB- and to subsequent protonation of a doubly reduced plastoquinone QB2-.

Cyanobacteria↗

Interaction of carbonyl cyanide m-chlorophenylhydrazone with the photosystem II acceptor side.

We show that CCCP, known as an uncoupler of photophosphorylation and an ADRY agent, inhibits FeCy photoreduction and coupled O2 evolution by isolated chloroplasts equally (I50 approximately 2 microM), but is practically without effect on the O2 evolution coupled with SiMo reduction within the 0.2-10 microM concentration range. CCCP has no effect on the nanosecond chlorophyll fluorescence in chloroplasts incubated at low light intensity, but decreases it at high light intensity. The electron transfer from reduced TMPD or duroquinol to methylviologen is resistant to CCCP. The efficiency of the CCCP inhibitory action on the FeCy photoreduction depends on the rate of electron flow, which is controlled by the light intensity. The data obtained show that CCCP is oxidized by the photosystem II donor side and is reduced by QP, competing for electrons with FeCy and the cytochrome blf complex.

Carbonyl Cyanide m-Chlorophenyl Hydrazone↗

Technical bioenergetics and ecosystem biotechnology.

The present work explores some possible practical uses of bioenergetic processes (fermentation, respiration and photosynthesis) in the microbial cell for the purpose of renewable fuel production. These considerations are based on ecosystem biotechnology which include the following main points: 1. Creation of closed energy-transducing ecosystems under laboratory conditions. 2. Production of renewable energy carriers with the help of microbial communities (e.g., mixed cultures of microorganism). 3. Investigation of the role of the individual microoganism in the population or community.

Bacteria↗

[Dependence of Bacillus subtilis cell respiration on monovalent cations].

Nigericin, monensin, valinomycin + carbonyl-cyanide-m-chlorophenylhydrazone and gramicidin inhibit the respiration of Bacillus subtilis cells incubated with NAD-dependent substrates or succinate, but not with ascorbate + N,N,N',N'-tetramethyl-p- phenylene-diamine. The level of inhibition was decreased by potassium ions and, in a lower degree, by sodium or ammonium ions. The results obtained suggest that the respiration of Bacillus subtilis depends on the presence of monovalent cations whose effects seem to be directed at complexes I, III and probably complex II of the respiratory chain.

Bacillus subtilis↗

Effect of polyelectrolytes on serine proteinase secretion by Bacillus subtilis.

Addition of polycations with molecular masses of 5-40 kDa as well as Na+, stimulated serine proteinase secretion by Bacillus subtilis cells. Polyanions and higher-molecular-mass polycations (100-200 kDa) were inefficient. The enzyme yields in the presence of polycations or Na+ were equal in magnitude. The results indicate that the cations, apparently counteracting the negative surface charge of the bacterial plasma membrane, cause the desorption of the serine (alkaline) proteinase. The synthesis of the proteinase is inferred to be stopped as the enzyme is bound to the outer surface of the plasma membrane. The desorption of the enzyme thus induces the synthesis of the new portions of proteinase.

Bacillus subtilis↗

[The surface membrane charge of bacteria and its role in serine proteinase secretion by Bacillus subtilis cells].

Univalent, bivalent and trivalent metal cations increase the fluorescence yield of 9-aminoacridine in the suspensions of chromatophores of the purple nonsulfur bacterium Rhodospirillum rubrum isolated thylakoid membranes and cells of cyanobacterium Anabaena variabilis, cells Bacillus subtilis. The active cation concentrations increase about in 10 times with the decrease of their valency by one. It points to the fact that the changes in 9-aminoacridine fluorescence serve for the monitoring of the surface charge of bacterial membranes. The negative surface charge of B. subtilis cells increases before the onset of the serine protease secretion. The metal cations stimulate the serine protease secretion by B. subtilis cells, the stimulating effect correlates with the action of cations on the 9-aminoacridine fluorescence yield. It is suggested that the surface charge of cytoplasmic membrane regulates the formation and release of serine protease by the cells of B. subtilis.

Aminacrine↗

[Quinones and their interactions with enzyme complexes of energy-transducing biomembranes].

The functionally essential properties of biomembrane quinones and the mechanism of their interaction with protein components are discussed. The hypotheses on the mobile quinone pool or the ability of protein-bound quinones to transfer redox equivalents in biomembranes are discussed. The idea of quinone domains is invoked, and evidence is provided for the presence of such domains in operative biomembranes.

Animals↗

The dibromothymoquinone effect on membrane potential generation in Rhodospirillum rubrum chromatophores.

2,5-Dibromo-3-methyl-6-isopropyl benzoquinone (DBMIB) inhibits the light-dependent membrane potential generation in Rhodospirillum rubrum chromatophores. The inhibition is relieved by electron donors and is obviously due to oxidation of the photosynthetic electron transfer chain components. In addition, high DBMIB concentrations elicit another effect probably caused by disruption of quinone functions in chromatophores. However, in quinone-depleted chromatophores and proteoliposomes containing the P-870 reaction center and light-harvesting antenna complexes, DBMIB stimulates membrane potential generation in the light, probably restoring some of the quinone-dependent processes in the membrane. DBMIB inhibits the inorganic pyrophosphate- and ATP-induced membrane potential generation in chromatophores.

Adenosine Triphosphate↗

[Effect of 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone, a ubiquinone analog, and SH-reagents on the electrogenic function of pyrophosphatase from Rhodospirillum rubrum chromatophores].

Inorganic pyrophosphate-induced membrane potential generation in Rhodospirillum rubrum chromatophores is inhibited by 2,5-dibromo-2-methyl-6-isopropyl-p-benzoquinone (DBMIB). The inhibition is increased by menadione and dithionite and is not relieved by ascorbate; this effect is probably due to a displacement of the intrachromatophore quinones by DBMIB. The SH-reagents, N-ethylmaleimide and p-chloromercurybenzoate at high concentrations inhibit membrane potential generation driven by inorganic pyrophosphate hydrolysis, thus suggesting the involvement of SH-groups in pyrophosphatase operation. The mechanism of regulation of the mode of action of pyrophosphatase by quinones and the location of the SH-groups of the enzyme are discussed.

Bacterial Chromatophores↗

[Dicyclohexylcarbodiimide as an inhibitor of light- and pyrophosphate-induced formation of membrane potential in chromatophores of purple bacteria].

N,N'-Dicyclohexylcarbodiimide (DCCD) suppresses the uptake of penetrating tetraphenylborate anions by Rhodospirillum rubrum chromatophores during cyclic and non-cyclic electron transfer and ATP and PP i hydrolyses. The photochemical activity of the bacteriochlorophyll reaction centers of the chromatophores in insensitive to DCCD. This supports the view that DCCD inhibits the electron transfer between the primary and secondary quinones of the photosynthetic chain. Incorporation of the chromatophores into a planar phospholipid-decane membrane abolishes or considerably reduces the inhibiting effect of DCCD on the membrane potential generation during the light-induced electron transfer and PP i (but not ATP) hydrolysis. The inhibition of the photosynthetic electron transfer is proposed to be due to the effect of DCCD as a quinone antagonist which competes with the secondary quinone for the binding at the active site. By expelling quinones DCCD seems to destroy the specific microenvironment of PPiase in the membrane and to inhibit its catalytic activity. In the system with the planar membrane decane and/or phospholipids remove the effect of DCCD as a quinone antagonist.

Bacterial Chromatophores↗

[Generation of the differences of the electric potentials by Rhodospirillum rubrum reaction center complexes devoid of the heavy subunit].

The electrogenic activity of Rhodospirillum rubrum P870 reaction center complexes devoid of the heavy (H) subunit and retaining the light (L) and medium (M) subunits, was studied. The proteoliposomes containing such reaction center complexes were formed by a self-assembly procedure, using soya bean phospholipids. In the presence of Ca2+ the reaction center proteoliposomes were incorporated into a phospholipid-impregnated Teflon filter separating two solutions of identical composition. After addition of N,N,N',N'-tetra-methyl-p-phenylenediamine (or cytochrome c) and qinone (menadione), illumination caused generation of an electric potential difference between the two filter-separated compartments, the proteoliposome-free compartment being negatively charged. The illuminated proteoliposomes took up penetrating tetraphenylphosphonium cations and, in a lesser degree, tetraphenylborate anions. The data obtained suggest that the reaction center complexes containing only L- and M-subunits possess the electrogenic activity. The H-subunit is not directly involved in membrane potential generation.

Bacterial Proteins↗

The inhibition of photosynthetic electron transfer in Rhodospirillum rubrum by N ,N' -dicyclohexylcarbodiimide.

N ,N' -Dicyclohexylcarbodiimide (DCCD) at concentrations above 0.1 mM inhibits light-induced generation of a membrane potential in the course of cyclic and non-cyclic electron transfer, as well as light-induced oxygen uptake due to interaction of photoreduced secondary (loosely bound) ubiquinone with O2 in Rhodospirillum rubrum chromatophores. Similarly to o-phenanthroline, DCCD blocks the electron transfer in the chromatophores between the primary (tightly bound) and secondary ubiquinones.

Carbodiimides↗

Reconstitution of biological molecular generators of electric current. Inorganic pyrophosphatase.

Proteoliposomes have been reconstituted from soy-bean phospholipids (asolectin) and inorganic pyrophosphatase isolated from Rhodospirillum rubrum chromatophores. In the presence of Mg2+ ions, pyrophosphatase proteoliposomes were incorporated into a phospholipid-impregnated Teflon filter separating two solutions of an identical electrolyte content. Addition of inorganic pyrophosphate to the same compartment as proteoliposomes was found to induce generation of an electric potential difference between the two filter-separated compartments, the proteoliposomes-containing compartment being negatively charged. An electric potential difference of 15 mV and a current of 20 pA were observed. The electrogenic effect required Mg2+ and proved to be sensitive to fluoride, an inorganic pyrophosphatase inhibitor. Treatment with 10 microM N,N'-dicyclohexylcarbodiimide for several minutes was without influence upon pyrophosphate-induced membrane potential generation. Similar results were obtained in experiments with a proteoliposome suspension and a penetrating anion, tetraphenyl borate, which is a probe for membrane potential. The obtained data are discussed in connection with the results of studies on other enzymes as molecular generators of electric current.

Bacterial Chromatophores↗

[Cyclic electron transfer and membrane potential generation in chromatophores on non-sulfur bacteria Rhodospirillum rubrum].

The uptake of permeant anions by cells and chromatophores of the non-sulfur purple bacteria R. rubrum has been studied. Antimycin A causes biphasic inhibition of the light-induced uptake of tetraphenylborate anions (TB-) by the cells and the isolated chromatophores incubated under anaerobic conditions. The first phase is observed at small concentrations of antimycin and is due to its effect as an inhibitor of the cyclic electron transfer. The second phase is observed at concentrations higher than 1 microM and is due to its effect as an uncoupler of photophosphorylation. The inhibitory effect of antimycin is greatly enhanced under aerobic conditions and is due to its effect as an uncoupler of photophosphorylation. The innic cyclic redox chain in the isolated and intracellular chromatophores is apparently operated in two regimens: 1) as a chain including all redox components and, 2) as a chain functioning without cytochromes of the b type.

Antimycin A↗

[Structural organization of membranes reconstituted from phospholipids and subchromatophore pigment-protein complexes].

Pigment--protein complexes of the P870 reaction centers and complexes of the bacteriochlorophyll light-harvesting antenna were isolated from the chromatophores of the non-sulfur purple bacterium Rhodospirillum rubrum by solubilization with detergents. The proteoliposomes containing the reaction centers or reaction centers and the light-harvesting antenna as well as liposomes formed from phospholipids were obtained by a self-assembly procedure using seya bean phospholipids. The freeze-fracture study showed that the proteoliposomes contain a large amount of globular particles. The particles incorporated into the two types of the proteoliposomes were distinguished in size. The globules of the reaction center and antenna complexes were bigger in size than the reaction center globules. The globular structures were not found in the liposomal membranes. The liposomes formed in the absence of the pigment--protein complexes were predominantly the multilamellar vesicles. The proteoliposomes were mostly represented as monolamellar membrane vesicles. The spatial arrangement of the reaction center complexes in the membranes is discussed.

Bacterial Chromatophores↗

Blue and red shifts of bacteriochlorophyll absorption band around 880 nm in Rhodospirillum rubrum.

The redox potential dependence of the light-induced absorption changes of bacteriochlorophyll in chromatophores and subchromatophore pigment-protein complexes from Rhodospirillum rubrum has been examined. The highest values of the absorption changes due to the bleaching of P-870 and the blue shift of P-800 in chromatophores and subchromatophore complexes are observed in the 360-410mV redox potential range. At potentials below 300 mV (pH 7.0), the 880 nm band of bacteriochlorophyll shifts to shorter wavelengths in subchromatophore complexes and to longer wavelengths in chromatophores. The data on redox titration show that the red and blue shifts of 880-nm bacteriochlorophyll band represent the action of a non-identified component (C340) which has an oxidation-reduction midpoint potential close to 340 mV (n=1) at pH 6.0--7.6. The Em of this component varies by 60 mV/pH unit between pH 7.6 and 9.2. The results suggest that the red shift is due to the transmembrane, and the blue shift to the local intramembrane electrical field. The generation of both the transmembrane and local electrical fields is apparently governed by redox transitions of the component C340.

Bacterial Chromatophores↗