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Biomedical subjects

V D Samuilov

Publications and source records attributed to V D Samuilov.

At least 55 records · Page 3Linked to original sources

Two regimens of electrogenic cyclic redox chain operation in chromatophores of non-sulfur purple bacteria. A study using antimycin A.

Antimycin A causes a biphasic suppression of the light-induced membrane potential generation in Rhodospirillum rubrum and Rhodopseudomonas sphaeroides chromatophores incubated anerobically. The first phase is observed at low antibiotic concentrations and is apparently due to its action as a cyclic electron transfer inhibitor. The second phase is manifested at concentrations which are greater than 1--2 muM and is due to uncoupling that may be connected with an antibiotic-induced dissipation of the electrochemical H+ gradient across the chromatophore membrane. The inhibitory effect of antimycin added at low concentrations under aerobic conditions is removed by succinate to a large extent. It is expected that the electrogenic cyclic redox chain in the bacterial chromatophores incubed under conditions of continuous illumination may function at two regimes: (1) as a complete chain involving all the redox components, and (2) as a shortened chain involving only the P-870 photoreaction center, ubiquinone and cytochrome c2.

Anaerobiosis↗

[Reconstitution of electrogenic function of pyrophosphatase isolated from Rhodospirillum rubrum membranes].

The membrane vesicles (proteoliposomes) have been reconstituted from phospholipids and inorganic pyrophosphatase (EC 3.6.1.1) isolated from Rhodospirillum rubrum chromatophores. An addition of inorganic pyrophosphate (PPi) causes a Mg2+-dependent formation of a transmembrane electric potential difference and an uptake of penetrating tetraphenylborate anions by the proteoliposomes. Thus, isolated pyrophosphatase, being incorporated into the phospholipid membrane, functions as a MgPPi-dependent protein generator of the electric current.

Cell Membrane↗

[Shifts of the bacteriochlorophyll absorption band at 880 nm in chromatophores and subchromatophore pigment-protein complexes from Rhodospirillum rubrum].

The redox potential dependency of the light-induced absorption changes of bacteriochlorophyll in the chromatophores and subchromatophore particles from Rhodospirillum rubrum has been studied. The highest values of the absorption changes due to the bleaching of P870 and the blue shift of P800 are observed within the redox potential range of 360--410. At the potential values below 300 mV the 880 nm band of bacteriochlorophyll shifts to shorter wavelengths in the subchromatophore particles and to longer wavelengths in the chromatophores. Redox titration revealed that the red and blue shifts of 880 nm bacteriochlorophyll band are caused by the functioning of a non-identified component (X) which has an oxidation -- reduction midpoint potential close to 340 mV (n = 1) within the pH range of 6,0--7,6. The Em for this component decreases by 60 mV/pH unit within the pH range of 7.6--9,2. The results obtained suggest that the red shift is due to the transmembrane, while the blue shift -- to the local intramembrane electric field. The generation of both the transmembrane and local intramembrane electric fields apparently depends on redox transitions of the component X.

Bacterial Chromatophores↗

[Light-induced oxygen uptake by chromatophores and subchromatophore pigment-protein complexes of Rhodospirillum rubrum].

Chromatophores of R. rubrum incubated with electron donors, e. g. reduced diaminodurene, TMPD, phenazine methosulphate, cytochrome c or ferrocyanide, are able to catalyze O2 uptake upon illumination. This process is inhibited by o-phenanthroline as well as upon extraction of quinones from chromatophores, but not by antimycin A, rotenone or CN-. The O2 uptake sensitive to the action of o-phenanthroline is also observed in the illuminated subchromatophore P870 reaction center complexes and reaction center plus light-harvesting antenna complexes incubated with electron donors, quinones and detergents. The data obtained are in agreement with a suggestion that the photooxidase activity of chromatophores and subchromatophore pigment-protein complexes is due to the interaction of photoreduced ubiquinone with O2.

Bacterial Chromatophores↗

Reconstitution of biological molecular generators of electric current. Bacteriochlorophyll and plant chlorophyll complexes.

1. Electric generation by bacteriochlorophyll reaction center complexes from Rhodospirillum rubrum and by photosystem I complexes from pea chloroplasts has been studied. 2. The methods for the proteoliposome reconstitution from azolectin and bacteriochlorophyll- or plant chlorophyll-containing protein complexes have been elaborated. Light-dependent electric responses of the proteoliposomes were detected using (a) phenyldicarbaundecarborane anion (PCB-) probe and (b) direct measurement by a voltmeter in the proteoliposome-planar phospholipid membrane system. 3. Both PCB- and direct measurements demonstrated that bacteriochlorophyll proteoliposomes are competent in light-dependent electric generation (plus outside proteoliposomes). The photoelectric effect was shown to increase on addition of tetramethyl-p-phenylenediamine (TMPD), CoQ6, and vitamin K3, and to decrease on addition of ferricyanide, o-phenanthroline and a protonophorous uncoupler. Estimation of the photoelectromotive force of the bacteriochlorophyll proteoliposome-planar membrane system gave a value of about 0.2 V. The action spectrum of the photoelectric effect was similar to the absorption spectrum of the bacteriochlorophyll complex. 4. Reconstitution of proteoliposomes containing bacteriochlorophyll centers and bacteriorhodopsin resulted in the system generating an electric field whose direction can be changed by varying the spectral composition of the light: the red light, exciting bacteriochlorophyll, induces negative, and the green light, exciting bacteriorhodopsin, induces positive charging of the proteoliposome interior. 5. Association of isolated R. rubrum chromatophores with planar phospholipid membrane was found to give a system demonstrating light-induced electric generation as high as 215 mV in the presence of napthoquinone, TMPD (or phenazine methosulfate, PMS), and ascorbate. Under the same conditions, addition of inorganic pyrophosphate or ATP results in formation of an electric field of the same direction as that induced by light. 6. Proteoliposomes with plant chlorophyll complexes of photosystem I demonstrated light-induced PCB- responses indicating formation of the electric field with plus inside vesicles. The effect required PMS addition. A protonophorous uncoupler and o-phenanthroline were inhibitory. Electric responses in the chlorophyll proteoliposome-planar membrane system were very small (not higher than 10 mV).

Bacterial Chromatophores↗

Generation of electric current by chromatophores of Rhodospirillum rubrum and reconstitution of electrogenic function in subchromatophore pigment-protein complexes.

Lipoprotein complexes, containing (1) bacteriochlorophyll reaction centers, (2) bacteriochlorophyll light-harvesting antenna or (3) both reaction centers and antenna, have been isolated from chromatophores of non-sulphur purple bacteria Rhodospirillum rubrum by detergent treatments. The method of reconstituting the proteoliposomes containing these complexes is described. Being associtated with planas azolectin membrane, ptoteoliposomes as well as intact chromatophores were found to generate a light-dependent transmembrane electric potential difference measured by Ag/AgC1 electrodes and voltmeter. The direction of the electric field inproteoliposomes can be regulated by the addition of antenna complexes to the reconstitution mixture. The reaction center complex proteoliposomes generate an electric field of a direction opposite to that in chromatophores, whereas proteoliposomes containing reaction center complexes and a sufficient amount of antenna complexes produce a potential difference as in chromatophores. ATP and inorganic pyrophosphate, besides light, were shown to be usable as energy sources for electric generation in chromatophores associated with planar membrane.

Bacterial Chromatophores↗

[Reconstruction of the function of membrane potential formation by isolated pigment-protein complexes of Rhodospirillum rubrum].

The pigment-protein complexes, containing the photosynthetic reaction centers and chlorophyll antenna, were isolated from non-sulfur purple bacteria Rhodospirillum rubrum, by use of cholate. Absorption spectra and quantum yield of photo-oxidation of bacteriochlorophyll P870 of complexes were similar to those of chromatophores. The procedure of reconstitution of proteoliposomes from phospholipids and complexes was proposed. The proteoliposomes incubated with CoQ6 and TMPD upon illumination are generating the transmembrane electric potential difference (plus inside proteoliposomes). The electric potential difference was registered by three independent methods: by transmembrane electrophoresis of penetrating anions PCB-, by direct measurement with using voltmeter in system "proteoliposome--planar membrane" and electrochromic absorption band shifts of bacteriochlorophyll.

Chemical Phenomena↗

A study on the membrane potential and pH gradient in chromatophores and intact cells of photosynthetic bacteria.

Generation of membrane potential (delta psi) and transmembrane pH difference (delta pH) was studied in PPi-energized chromatophores of Rhodospirillum rubrum by means of measurements of carotenoid and bacteriochlorophyll absorption changes, atebrin and 8-anilinonaphthalene-1-sulphonate fluorescence responses, and phenyldicarbaundecaborane transport. The data obtained are consistent with the suggestion that carotenoid, bacteriochlorophyll and phenyldicarbaundecaborane responses are indicators of delta psi, while an atebrin response is an indicator of delta pH. The fluorescence of 8-anilinonaphthalene-1-sulphonate is affected both by delta psi and delta pH.

Anilino Naphthalenesulfonates↗