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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : BONDAR V.S., TROFIMOV K.P., SANDALOV T.P., VYSOTSKII E.S.
Заглавие : EFFECT OF TEMPERATURE ON ACTIVITY AND STABILITY OF OBELIN
Место публикации : Biochem.-Moscow: PLENUM PUBL CORP, 1992. - Vol. 57, Is. 7. - С. 717-724. - 8. - ISSN 0006-2979
Примечания : Cited References: 15
Предметные рубрики: CA-2+
PHOTOPROTEINS
INDICATORS
Ключевые слова (''Своб.индексиров.''): photoproteins--obelin--activation energy--thermoinactivation--thermostability
Аннотация: The temperature dependence of bioluminescent activity of the Ca2+-activated photoprotein obelin from the hydroid polyp Obelia longissima and thermoinactivation of this protein at different concentrations of (NH4)2SO4 have been studied. The maximal intensity of luminescence of obelin was observed at 4-15-degrees-C. The activity of the photoprotein is completely stable to storage for 3 days at room temperature. Increasing the temperature to 40-degrees-C resulted in a 25-30% loss of enzyme activity in 1 h. The presence of ammonium sulfate during heating stabilizes the activity of obelin. Two breaks, at 11 +/- 3-degrees-C and 47 +/- 3-degrees-C, are observed in the Arrhenius plot of the first-order rate constant of the luminescence decay. The bioluminescent curves of obelin are biphasic in the temperature range 10-40-degrees-C. It is assumed that obelin may exist in two kinetically distinct conformers (active and inactive) whose ratio is temperature dependent.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : MATVEEV S.V., ILLARIONOV B.A., VYSOTSKI E.S., BONDAR V.S., MARKOVA S.V., ALAKHOV Y.B.
Заглавие : OBELIN MESSENGER-RNA - A NEW TOOL FOR STUDIES OF TRANSLATION IN CELL-FREE SYSTEMS
Колич.характеристики :6 с
Место публикации : Anal. Biochem.: ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS, 1995. - Vol. 231, Is. 1. - С. 34-39. - ISSN 0003-2697, DOI 10.1006/abio.1995.1499
Примечания : Cited References: 17
Предметные рубрики: MESSENGER-RNA
AEQUORIN
PROTEIN
CLONING
CDNA
Аннотация: Obelin mRNA obtained in vitro with the aid of SP6 RNA polymerase was translated in a wheat germ cell-free system, Only the polypeptide with a molecular mass of about 20 kDa was synthesized. The activation of apoobelin with a synthetic coelenterazine revealed a luminescence activity initiated by calcium. The specific activity was 3.6 +/- 0.4 x 10(15) photons per mg of the in vitro synthesized obelin (k = 6.9 s(-1)). The luminescence of the obelin was in a good correlation with the protein concentration calculated by the incorporation of [C-14]Leu. The determination of the amount of de novo synthesized obelin based on measurement of its luminescence is one-thousand times more sensitive than the approach based on the incorporation of labeled amino acid. Thus, obelin mRNA has some advantages for evaluating the efficiency of cell-free translation when compared with standard methods. (C) 1995 Academic Press, Inc.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Setkov N.A.
Заглавие : Negative control of cell proliferation depend on protein synthesis in resting cells
Колич.характеристики :4 с
Место публикации : Dokl. Akad. Nauk: MEZHDUNARODNAYA KNIGA, 1996. - Vol. 346, Is. 5. - P708-711. - ISSN 0869-5652
Примечания : Cited References: 15
Предметные рубрики: C-FOS
SYNTHESIS INHIBITORS
3T3 CELLS
TRANSCRIPTIONAL ACTIVATION
DNA-SYNTHESIS
FIBROBLASTS
NUCLEI
FUSION
MYC
JUN
WOS
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Tyulkova N.A., Sandalova T.P.
Заглавие : Comparative study of temperature effects on bacterial luciferases
Колич.характеристики :10 с
Место публикации : Biochem.-Moscow: PLENUM PUBL CORP, 1996. - Vol. 61, Is. 2. - P205-214. - ISSN 0006-2979
Примечания : Cited References: 23
Предметные рубрики: BIOLUMINESCENCE
Ключевые слова (''Своб.индексиров.''): bacterial luciferase--temperature--activation energy
Аннотация: Effects of temperature on bioluminescent patterns of luciferases from luminescent bacteria Vibrio harveyi, Vibrio fischeri, Photobacterium leiognathi, and Photobacterium phosphoreum were studied. The highest luminescence level was observed at 15-25 degrees C for the luciferase from P. phosphoreum, at 20-30 degrees C for the V. fischeri and P. leiognathi enzymes, and at 30-37 degrees C for the enzyme from V. harveyi. All the luciferases were significantly stabilized at increased salt concentrations, at low pH values, or in the presence of dithiothreitol (DTT) and EDTA. The addition of DTT and EDTA affected the reversible stage of enzyme inactivation, while salts reduced the rate of the irreversible stage. A peak corresponding to aggregated protein was detected by gel chromatography of irreversibly inactivated luciferase. Activation energies were calculated for each luciferase in bioluminescent reactions with decanal, dodecanal, tetradecanal, and without aldehydes. The activation energy of the reaction with tetradecanal was much lower than those with the other aldehydes. The temperature dependence of the lifetime of the long-lived reaction intermediate showed that in the 10-30 degrees C interval all the luciferases, except for the enzyme from V. harveyi, have only one active form.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Kydryasheva N.S., Zuzikova E.V., Gutnik T.V.
Заглавие : Mechanism of action of metal salts on bacterial bioluminescent system in vitro
Колич.характеристики :7 с
Место публикации : Biofizika: MEZHDUNARODNAYA KNIGA, 1999. - Vol. 44, Is. 2. - P244-250. - ISSN 0006-3029
Примечания : Cited References: 21
Ключевые слова (''Своб.индексиров.''): bacterial bioluminescent system--metal salts, mechanism of action
Аннотация: The action of met al salts on the luciferase-NADH: FMN-oxydoheductase bacterial luminescent system in vitro was studied. The effect is shown to depend on the nature of cations rather than anions, The type of the effect (activation, inhibition, and absence of the effect) is determined by the structure of the; cation, the nature and occupancy of its external orbitals, and charge. The results were interpreted on the basis of the hypothesis that upper electron excited states of the emitter are involved in the bioluminescence process. Based on thermodynamic calculations, the type of the effect was related to the energetic characteristics of the cation, Presumably, the mechanism of action of cations is determined by their involvement in the formation and deactivation of electron excited states in the bioluminescence process.
WOS
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Karabutov A.V., Frolov V.D., Konov V.I., Ralchenko V.G., Gordeev S.K., Belobrov P.I.
Заглавие : Low-field electron emission of diamond/pyrocarbon composites
Колич.характеристики :6 с
Место публикации : J. Vac. Sci. Technol. B: A V S AMER INST PHYSICS, 2001. - Vol. 19: 13th International Vacuum Microlectronics Conference (AUG 13-17, 2000, GUANGZHOU, PEOPLES R CHINA), Is. 3. - P965-970. - ISSN 1071-1023, DOI 10.1116/1.1368669
Примечания : Cited References: 31
Предметные рубрики: CVD DIAMOND FILMS
SCANNING-TUNNELING-MICROSCOPY
AMORPHOUS-CARBON
COLD-CATHODE
Аннотация: Properties of the field electron emission for diamond/pyrocarbon nanocomposites produced from diamond particles surrounded by an sp(2)-bonded pyrocarbon matrix are considered as functions of a size of diamond particles selected in the range of 5 nm - 5 mum, and of an average thickness of the pyrocarbon shell controlled by the pyrocarbon/diamond mass ratio varied from 0 to 0.5. The low-threshold emission at fields of greater than or equal to1 V/mum with ''no activation/no hysteresis'' I-V behavior was observed for these materials using tungsten tip microprobes as well as a fluorescent screen. A specially designed scanning tunneling-field emission microscope was used for simultaneous mapping of field emission intensity, topography, work function, and electrical resistivity to study the mechanisms of the emission from the composites and well-emitting chemical vapor deposition diamond films. It was found that for both of the materials emission centers are associated with interfaces between diamond and sp2-bonded carbon phases. Possible mechanisms of the low-field electron emission for the diamond/graphite composites including local field enhancement are analyzed. A model of the low-field emission based on quantum well effect at the diamond/graphite interface is proposed and discussed. (C) 2001 American Vacuum Society.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Karabutov A.V., Frolov V.D., Konov V.I., Ralchenko V.G., Gordeev S.K., Belobrov P.I.
Заглавие : Low-field electron emission of diamond/pyrocarbon composites
Место публикации : Journal of Vacuum Science and Technology B: Microelectronics and Nanometer Structures. - 2001. - Vol. 19: 13th International Vaccum Microelectronics Conference (14 August 2000 through 17 August 2000, Guangzhou, Is. 3. - С. 965-970. - ISSN 10711023 (ISSN) , DOI 10.1116/1.1368669
Ключевые слова (''Своб.индексиров.''): carbon nanotubes--chemical bonds--chemical vapor deposition--composite materials--diamond films--electric conductivity--electron emission--electron energy levels--hysteresis--interfaces (materials)--raman scattering--semiconducting diamonds--semiconductor quantum wells--transmission electron microscopy--x ray diffraction analysis--x ray photoelectron spectroscopy--pyrocarbon composites--nanostructured materials
Аннотация: The properties of field electron emission for diamond/pyrocarbon nanocomposites produced from diamond particles surrounded by a pyrocarbon matrix were studied. Low-threshold emissions at fields of ?1 V/?m with no activation or hysterisis in the current versus voltage (I/V) behaviour were observed for the materials. Scanning tunneling-field emission microscopy was used to study the mechanisms of low-field electron emission from the composites, and a model based on quantum well effect at the diamond/graphite interface was proposed and discussed.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Gerasimova M.A., Kudryasheva N.S.
Заглавие : Effects of potassium halides on bacterial bioluminescence
Колич.характеристики :5 с
Место публикации : J. Photochem. Photobiol. B-Biol.: ELSEVIER SCIENCE SA, 2002. - Vol. 66, Is. 3. - P218-222. - ISSN 1011-1344, DOI 10.1016/S1011-1344(02)00240-3
Примечания : Cited References: 20
Предметные рубрики: COMPONENTS
SEWAGE
Ключевые слова (''Своб.индексиров.''): bioluminescence--halides--heavy atom--inhibition--activation
Аннотация: The effects of potassium halides KCl, KBr and KI on NADH:FMN-oxidoreductase-luciferase bioluminescent coupled enzyme system were studied. The influence of salt additions on bioluminescence intensity and bioluminescence light yield was investigated. The inhibition and activation parameters of the salts were calculated using their dependencies on concentration of the salts. The correlation between the inhibition of bioluminescence intensity and the halide mass was demonstrated: the inhibiting ability of the salts increases with the increase of atomic weight of the anions. The inhibition parameters increase and the activation parameters decrease, accordingly. (C) 2002 Elsevier Science B.V. All rights reserved.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Bolsunovskij A.Ya., Ermakov A.I., Burger M., Degermendzhy A.G., Sobolev A.I.
Заглавие : Accumulation of artificial radionuclides by the Yenisei river aquatic plants in the area affected by the activity of the Mining-and-Chemical Combine
Место публикации : Radiatsionnaya Biologiya. Radioekologiya. - 2002. - Vol. 42, Is. 2. - С. 194-199. - ISSN 08698031 (ISSN)
Ключевые слова (''Своб.индексиров.''): contamination--ecosystems--environmental impact--gamma ray spectrometers--radioisotopes--river pollution--radioecological concentration--radioisotope accumulation--the yenisei river--ecology--plutonium--radioisotope--strontium--article--chemical industry--chemistry--comparative study--gamma spectrometry--industrial waste--mining--nuclear reactor--plant--radiochemistry--russian federation--water pollutant--chemical industry--industrial waste--mining--nuclear reactors--plants--plutonium--radiochemistry--radioisotopes--siberia--spectrometry, gamma--strontium radioisotopes--water pollutants, radioactive
Аннотация: The aim of the paper is to investigate accumulation of artificial radionuclides by the Yenisei river aquatic plants collected in the area affected by the activity of the Mining-and-Chemical Combine (Zheleznogorsk) from 1997 to 2000. The samples of aquatic plants were of four species: Potamogeton lucens, Fontinalis antipyretica, Elodea canadensis, and Ceratophyllum demersum. The gamma-spectrometric analysis of the samples of aquatic plants for artificial radionuclides revealed a wide spectrum of radionuclides. Radionuclides of activation origin were found in the aquatic plants taken both near the Combine and 200 km down of it. The radiochemical analysis of aquatic plants revealed strontium and plutonium isotopes. Among the aquatic plants, the highest concentration factors for the principal radionuclides were recorded in Fontinalis antipyretica (water moss).
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10.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Sevastianov V.I., Perova N.V., Shishatskaya E.I., Kalacheva G.S., Volova T.G.
Заглавие : Production of purified polyhydroxyalkanoates (PHAs) for applications in contact with blood
Колич.характеристики :14 с
Место публикации : J. Biomater. Sci.-Polym. Ed.: VSP BV, 2003. - Vol. 14, Is. 10. - P1029-1042. - ISSN 0920-5063, DOI 10.1163/156856203769231547
Примечания : Cited References: 34
Предметные рубрики: BIODEGRADABLE POLYESTERS
POLYMERS
Ключевые слова (''Своб.индексиров.''): polyhydroxyalkanoates (phas)--polyhydroxybutyrate (phb)--poly(hydroxybutyrate-co-hydroxyvalerate) (phbv)--endotoxins--beta-hydroxy acids--hemocompatibility
Аннотация: Samples of olyhydroxyalkanoates (PHAs), polyhydroxybutyrate (PHB) and copolymers poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) with 4 and 18 mol% hydroxyvalerate, synthesized by the bacteria Ralstonia eutropha B5786, were investigated. PHA films in contact with blood did not activate the hemostasis system at the level of cell response, but they did activate the coagulation system and the complement reaction. To detect biologically-active components in the PHAs, a detailed analysis of the composition of the polymers was conducted. Gas chromatography-mass spectrometry revealed long-chain fatty acids (FAs) in the tested PHAs. Their total concentration in the polymer ranged from tenths of mol% to 2-3 mol%, depending on the purification method. C-16:0 constituted the largest proportion, up to 70%. Of the long-chain hydroxy acids, only beta-OH-C-14:0 was detected and it did not exceed 0.06 mol%. The analysis of the hemocompatibility properties of the PHAs purified by a specialized procedure, including the quantitative and morphological estimation of platelets adherent to the surface of polymer films, the plasma recalcification time and complement activation studies, indicated that PHB and PHBV can be used in contact with blood. It has been found out that the lipopolysaccharides of bacteria producing PHAs, which contain mostly long-chain hydroxy acids, can be the factor activating the hemostasis systems. Thus, the technology of PHA purification must satisfy rather stringent specific requirements.
WOS
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11.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Sevastianov V.I., Perova N.V., Shishatskaya E.I., Kalacheva G.S., Volova T.G.
Заглавие : Production of purified polyhydroxyalkanoates (PHAs) for applications in contact with blood
Место публикации : Journal of Biomaterials Science, Polymer Edition. - 2003. - Vol. 14, Is. 10. - С. 1029-1042. - ISSN 09205063 (ISSN) , DOI 10.1163/156856203769231547
Ключевые слова (''Своб.индексиров.''): ?-hydroxy acids--endotoxins--hemocompatibility--poly(hydroxybutyrate-co-hydroxyvalerate) (phbv)--polyhydroxyalkanoates (phas)--polyhydroxybutyrate (phb)--bacterium lipopolysaccharide--carbon--complement--copolymer--hydroxyacid--long chain fatty acid--poly(3 hydroxybutyric acid)--polyhydroxyalkanoic acid--valeric acid derivative--adult--article--biofilm--biotechnology--blood analysis--blood clotting--blood compatibility--cell function--chemical analysis--chemical composition--complement activation--concentration (parameters)--controlled study--gas chromatography--hemostasis--human--human cell--mass spectrometry--micromorphology--nonhuman--priority journal--purification--quantitative analysis--sampling--synthesis--thrombocyte adhesion--wautersia eutropha--biocompatible materials--blood--blood coagulation tests--chromatography, gas--complement activation--cupriavidus necator--fatty acids--humans--platelet adhesiveness--polyesters--surface properties
Аннотация: Samples of olyhydroxyalkanoates (PHAs), polyhydroxybutyrate (PHB) and copolymers poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) with 4 and 18 mol% hydroxyvalerate, synthesized by the bacteria Ralstonia eutropha B5786, were investigated. PHA films in contact with blood did not activate the hemostasis system at the level of cell response, but they did activate the coagulation system and the complement reaction. To detect biologically-active components in the PHAs, a detailed analysis of the composition of the polymers was conducted. Gas chromatography-mass spectrometry revealed long-chain fatty acids (FAs) in the tested PHAs. Their total concentration in the polymer ranged from tenths of mol% to 2-3 mol%, depending on the purification method. C16:0 constituted the largest proportion, up to 70%. Of the long-chain hydroxy acids, only ?-OH-C14:0 was detected and it did not exceed 0.06 mol%. The analysis of the hemocompatibility properties of the PHAs purified by a specialized procedure, including the quantitative and morphological estimation of platelets adherent to the surface of polymer films, the plasma recalcification time and complement activation studies, indicated that PHB and PHBV can be used in contact with blood. It has been found out that the lipopolysaccharides of bacteria producing PHAs, which contain mostly long-chain hydroxy acids, can be the factor activating the hemostasis systems. Thus, the technology of PHA purification must satisfy rather stringent specific requirements.
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12.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Bolsunovsky A., Zotina T., Bondareva L.
Заглавие : Accumulation and release of 241Am by a macrophyte of the Yenisei River (Elodea canadensis)
Место публикации : Journal of Environmental Radioactivity. - 2005. - Vol. 81, Is. 1. - С. 33-46. - ISSN 0265931X (ISSN) , DOI 10.1016/j.jenvrad.2004.10.012
Ключевые слова (''Своб.индексиров.''): 241am--accumulation--laboratory experiments--release--submerged plant elodea canadensis--yenisei river--activation analysis--biomass--concentration (process)--effluents--plutonium--rivers--aquatic plants--elodea plant--macrophytes--radioactive contamination--transuranium elements--americium 241--river water--americium--americium nitrate--bioaccumulation--biological uptake--macrophyte--pollutant source--radioactive pollution--river water--submerged vegetation--article--biomass--concentration (parameters)--environmental factor--fractionation--laboratory--macrophyte--radioactivity--river--sampling--adsorption--chemistry--hydrocharitaceae--methodology--physiology--plant--radiation exposure--radiation monitoring--russian federation--sediment--soil pollutant--time--tissue distribution--water pollutant--eastern hemisphere--eurasia--russian federation--world--yenisei river--elodea canadensis--adsorption--americium--biomass--chemical fractionation--geologic sediments--hydrocharitaceae--plant shoots--radiation monitoring--rivers--russia--soil pollutants, radioactive--time factors--tissue distribution--water pollutants, radioactive
Аннотация: The source of radioactive contamination of the Yenisei River floodplain, including contamination with transuranic elements, is the Mining-and-Chemical Combine of the Russian Ministry of Atomic Energy, which has for many years been producing weapons-grade plutonium. Transuranic elements have been detected not only in the soil and sediment of the river but also in the biomass of aquatic plants. This work is an investigation of accumulation and release of 241Am by a submerged macrophyte of the Yenisei River (Elodea canadensis) in laboratory experiments. In 2000-2003, laboratory experiments were carried out with biomass of E. canadensis Mich. and filtered river water. The samples were collected from the Yenisei River upstream of the discharge of the Combine's radioactive effluent. The experiments showed that 241Am is accumulated by Elodea biomass: the activity concentration of 241Am can reach 3280 В± 240 Bq/g, with the concentration factor for 241Am 16 600 В± 2200 l/kg. Results of chemical fractionation have proved that in the course of 241Am accumulation by Elodea biomass, 241Am tightly bound to biomass increases from 11% to 27% of the total 241Am in the biomass. Release of 241Am from the decaying Elodea biomass has been evaluated experimentally. By the end of the experiment (lasting up to 127 days), the Elodea plants had lost up to 65% of their initial 241Am activity and the rate of 241Am release into the water environment reached 23 Bq/day. В© 2004 Elsevier Ltd. All rights reserved.
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13.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Rodionova N.S., Petushkov V.N.
Заглавие : Effect of different salts and detergents on luciferin-luciferase luminescence of the enchytraeid Fridericia heliota
Место публикации : Journal of Photochemistry and Photobiology B: Biology. - 2006. - Vol. 83, Is. 2. - С. 123-128. - ISSN 10111344 (ISSN) , DOI 10.1016/j.jphotobiol.2005.12.014
Ключевые слова (''Своб.индексиров.''): atp--bioluminescence--earthworms--ions--luciferin-luciferase systems--triton x-100--adenosine triphosphate--anion--bromine--calcium ion--carbonic acid--cation--chloride--chromium derivative--detergent--dodecyl sulfate sodium--inorganic salt--iodine--iron derivative--luciferase--luciferin--magnesium ion--manganese--nitrate--phosphate--sulfate--sulfite--triton x 100--annelid worm--article--bioluminescence--concentration (parameters)--controlled study--enzyme activation--enzyme activity--enzyme inhibition--enzyme mechanism--in vitro study--nonhuman--priority journal--qualitative analysis--quantitative analysis--adenosine triphosphate--animals--cations, divalent--cations, monovalent--detergents--firefly luciferin--kinetics--luciferases--luminescence--metals--oligochaeta--photobiology--salts--annelida--clitellata--earthworms (sp.)--enchytraeidae--fridericia heliota--oligochaeta (metazoa)--pheretima sieboldi
Аннотация: The study addresses the effect produced by different inorganic salts and detergents (SDS, Triton X-100, the Tween series) on the ATP-dependent bioluminescent reaction catalyzed by the luciferase of the new earthworm species Fridericia heliota (Annelida: Clitellata: Oligochaeta: Enchytraeidae). It has been shown that the effect of divalent metal salts on luminescence is determined by the action of cations. Three of them - Mg2+, Mn2+ and Ca2+ - can stimulate luciferase activity at concentrations varying within a wide range, and Mn2+ can act as a 100%-effective substitute for Mg2+ in F. heliota luminescence reaction in vitro. The inhibitory effect of monovalent metal salts on luminescence is largely determined by the action of the anion part of the molecule. The effectiveness of the inhibitory effect of anions increases in the following order: {Mathematical expression}. Of the sodium salts, dodecyl sulfate, which is an anionic detergent, produces the strongest inhibitory effect on luciferase. On the contrary, nonionic detergents produce a stimulatory effect on the F. heliota luciferase. The action of the most effective of them - Triton X-100 - is determined by its ability to reduce the actual concentration of lipid inhibitors in the reaction mixture. В© 2006 Elsevier B.V. All rights reserved.
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14.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Rozhko T.V., Kudryasheva N.S., Kuznetsov A.M., Vydryakova G.A., Bondareva L.G., Bolsunovsky A.Y.
Заглавие : Effect of low-level alpha-radiation on bioluminescent assay systems of various complexity
Место публикации : Photochem. Photobiol. Sci.: ROYAL SOC CHEMISTRY, 2007. - Vol. 6, Is. 1. - С. 67-70. - 4. - ISSN 1474-905X, DOI 10.1039/b614162p
Примечания : Cited References: 52
Предметные рубрики: LOW-DOSE RADIATION
IRRADIATION
TOXICITY
QUINONES
HORMESIS
PHENOLS
Аннотация: This study addresses the effects of low-level alpha-radiation on bioluminescent assay systems of different levels of organization: in vivo and in vitro. Three bioluminescent assay systems are used: intact bacteria, lyophilized bacteria, and bioluminescent system of coupled enzyme reactions. Solutions of Am-241(NO3)(3) are used as a source of alpha-radiation. It has been shown that activation processes predominate in all the three bioluminescent assay systems subjected to short-term exposure (20-55 h) and inhibition processes in the systems subjected to longer-term exposure to radiation. It has been found that these effects are caused by the radiation component of Am-241(3+) impact. The intensity of the Am-241(3+) effect on the bioluminescent assay systems has been shown to depend on the Am-241(3+) concentration, level of organization and integrity of the bioluminescent assay system. The bioluminescent assay systems in vivo have been found to be highly sensitive to Am-241(3+) (up to 10(-17) M).
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15.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Eremeeva E.V., Natashin P.V., Song L..., Zhou Y.G., van Berkel WJH, Liu Z.J., Vysotski E.S.
Заглавие : Oxygen Activation of Apo-obelin-Coelenterazine Complex
Колич.характеристики :7 с
Место публикации : ChemBioChem: WILEY-V C H VERLAG GMBH, 2013. - Vol. 14, Is. 6. - С. 739-745. - ISSN 1439-4227, DOI 10.1002/cbic.201300002
Примечания : Cited References: 46. - The work was supported by grants from the RFBR 12-04-91153, and NSFC 31270795 and 31021062, by the Russian Federation Government Program "Measures to Attract Leading Scientists to Russian Educational Institutions" (grant 11.G34.31.0058), "Molecular and Cellular Biology" of RAS, President of the Russian Federation "Leading Science School" (grant 1044.2012.2). E.V.E. was supported by a Wageningen University Sandwich PhD Fellowship Program.
Предметные рубрики: GREEN-FLUORESCENT PROTEIN
JELLYFISH CLYTIA-GREGARIA
CRYSTAL-STRUCTURE
CA2+-DISCHARGED PHOTOPROTEIN
ANGSTROM RESOLUTION
RECOMBINANT OBELIN
MOLECULAR-OXYGEN
AEQUORIN
CALCIUM
BIOLUMINESCENCE
Ключевые слова (''Своб.индексиров.''): aequorin--coelenterazine--luminescence--photoprotein--protein folding
Аннотация: Ca2+-regulated photoproteins use a noncovalently bound 2-hydroperoxycoelenterazine ligand to emit light in response to Ca2+ binding. To better understand the mechanism of formation of active photoprotein from apoprotein, coelenterazine and molecular oxygen, we investigated the spectral properties of the anaerobic apo-obelincoelenterazine complex and the kinetics of its conversion into active photoprotein after exposure to air. Our studies suggest that coelenterazine bound within the anaerobic complex might be a mixture of N7-protonated and C2() anionic forms, and that oxygen shifts the equilibrium in favor of the C2() anion as a result of peroxy anion formation. Proton removal from N7 and further protonation of peroxy anion and the resulting formation of 2-hydroperoxycoelenterazine in obelin might occur with the assistance of His175. It is proposed that this conserved His residue might play a key role both in formation of active photoprotein and in Ca2+-triggering of the bioluminescence reaction.
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16.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Eremeeva E.V., Markova S.V., van Berkel WJH, Vysotski E.S.
Заглавие : Role of key residues of obelin in coelenterazine binding and conversion into 2-hydroperoxy adduct
Колич.характеристики :7 с
Коллективы : RFBR [12-04-00131]; Programs of the Government of Russian Federation "Measures to Attract Leading Scientists to Russian Educational Institutions" [11.G34.31.0058]; "Molecular and Cellular Biology" of RAS, President of Russian Federation "Leading science school" [3951.2012.4]; Wageningen University Sandwich PhD-Fellowship Program
Место публикации : J. Photochem. Photobiol. B-Biol.: ELSEVIER SCIENCE SA, 2013. - Vol. 127. - С. 133-139. - ISSN 1011-1344, DOI 10.1016/j.jphotobiol.2013.08.012
Примечания : Cited References: 65. - The work was supported by RFBR grant 12-04-00131, by the Programs of the Government of Russian Federation "Measures to Attract Leading Scientists to Russian Educational Institutions" (grant 11.G34.31.0058), "Molecular and Cellular Biology" of RAS, President of Russian Federation "Leading science school" (grant 3951.2012.4). E.V.E. was supported by Wageningen University Sandwich PhD-Fellowship Program.
Предметные рубрики: CA2+-REGULATED PHOTOPROTEINS
SEQUENCE-ANALYSIS
CRYSTAL-STRUCTURE
APO-OBELIN
CA2+-BINDING PHOTOPROTEIN
VIOLET BIOLUMINESCENCE
AEQUORIN REGENERATION
ANGSTROM RESOLUTION
RECOMBINANT OBELIN
MNEMIOPSIS-LEIDYI
Ключевые слова (''Своб.индексиров.''): bioluminescence--coelenterazine--obelin--aequorin--photoprotein
Аннотация: Bioluminescence of a variety of marine organisms is caused by monomeric Ca2+-regulated photoproteins, to which a peroxy-substituted coelenterazine, 2-hydroperoxycoelenterazine, is firmly bound. From the spatial structure the side chains of Tyr138, His175, Trp179, and Tyr190 of obelin are situated within the substrate-binding pocket at hydrogen bond distances with different atoms of the 2-hydroperoxycoelenterazine. Here we characterized several obelin mutants with substitutions of these residues regarding their bioluminescence, coelenterazine binding, and kinetics of active obelin formation. We demonstrate that Tyr138, His175, Trp179, and Tyr190 are all important for coelenterazine activation; substitution of any of these residues leads to significant decrease of the apparent reaction rate. The hydrogen bond network formed by Tyr138, Trp179 and Tyr190 participates in the proper positioning of coelenterazine in the active site and subsequent stabilization of the 2-hydroperoxy adduct of coelenterazine. His175 might serve as a proton shuttle during 2-hydroperoxycoelenterazine formation. (C) 2013 Elsevier B.V. All rights reserved.
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17.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Selivanova M.A., Rozhko T.V., Devyatlovskaya A.N., Kudryasheva N.S.
Заглавие : Comparison of chronic low-dose effects of alpha- and beta-emitting radionuclides on marine bacteria
Место публикации : Cent. Eur. J. Biol. - 2014. - Vol. 9, Is. 10. - С. 951-959. - ISSN 16443632 (ISSN) , DOI 10.2478/s11535-014-0331-0
Ключевые слова (''Своб.индексиров.''): am-241--hormesis--luminous bacteria--peroxides--radiotoxicity--tritium
Аннотация: Effects of Americium-241 (241Am), alpha-emitting radionuclide of high specific radioactivity, and tritium (3H), beta-emitting radionuclide, on luminous bacteria Photobacterium phosphoreum were compared. Bioluminescence intensity served as a marker of bacterial physiological activity. Three successive stages in the bioluminescence response to 241Am and 3H were found under conditions of lowdose irradiation: (1) absence of effects, (2) activation, and (3) inhibition. They were interpreted in terms of bacterial response to stressfactor as stress recognition, adaptive response/syndrome, and suppression of physiological function (i.e. radiation toxicity). Times of bioluminescence activation (TBA) and inhibition (TBI) were suggested as parameters to characterize hormesis and toxic stages in a course of chronic low-dose irradiation of the microorganisms. Values of TBA and TBI of 241Am were shorter than those of 3H, revealing higher impact of alpha-irradiation (as compared to beta-irradiation) under comparable radiation doses. Increases of peroxide concentration and NADH oxidation rates in 241Am aquatic solutions were demonstrated; these were not found in tritiated water. The results reveal a biological role of reactive oxygen species generated in water solutions as secondary products of the radioactive decay. The study provides a scientific basis for elaboration of bioluminescence-based assay to monitor radiotoxicity of alpha- and beta-emitting radionuclides in aquatic solutions. © 2014 Versita Warsaw and Springer-Verlag Wien.
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18.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Malikova N.P., Burakova L.P., Markova S.V., Vysotski E.S.
Заглавие : Characterization of hydromedusan Ca2+-regulated photoproteins as a tool for measurement of Ca2+concentration
Место публикации :. - 2014. - ISSN 16182642 (ISSN) , DOI 10.1007/s00216-014-7986-2
Ключевые слова (''Своб.индексиров.''): aequorin--calcium--clytin--coelenterazine--mitrocomin--obelin
Аннотация: Calcium ion is a ubiquitous intracellular messenger, performing this function in many eukaryotic cells. To understand calcium regulation mechanisms and how disturbances of these mechanisms are associated with disease states, it is necessary to measure calcium inside cells. Ca2+-regulated photoproteins have been successfully used for this purpose for many years. Here we report the results of comparative studies on the properties of recombinant aequorin from Aequorea victoria, recombinant obelins from Obelia geniculata and Obelia longissima, recombinant mitrocomin from Mitrocoma cellularia, and recombinant clytin from Clytia gregaria as intracellular calcium indicators in a set of identical in vitro and in vivo experiments. Although photoproteins reveal a high degree of identity of amino acid sequences and spatial structures, and, apparently, have a common mechanism for the bioluminescence reaction, they were found to differ in the Ca2+ concentration detection limit, the sensitivity of bioluminescence to Mg2+, and the rates of the rise of the luminescence signal with a sudden change of Ca2+ concentration. In addition, the bioluminescence activities of Chinese hamster ovary cells expressing wild-type photoproteins also differed. The light signals of cells expressing mitrocomin, for example, slightly exceeded the background, suggesting that mitrocomin may be hardly used to detect intracellular Ca2+ without modifications improving its properties. On the basis of experiments on the activation of endogenous P2Y2 receptor in Chinese hamster ovary cells by ATP, we suggest that wild-type aequorin and obelin from O. longissima are more suitable for calcium detection in cytoplasm, whereas clytin and obelin from O. geniculata can be used for calcium measurement in cell compartments with high Ca2+ concentration. [Figure not available: see fulltext.] © 2014 Springer-Verlag Berlin Heidelberg.
Scopus
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19.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Malikova N.P., Burakova L.P., Markova S.V., Vysotski E.S.
Заглавие : Characterization of hydromedusan Ca2+-regulated photoproteins as a tool for measurement of Ca(2+)concentration
Колич.характеристики :12 с
Коллективы : RFBR [12-04-00131]; Government of the Russian Federation [11.G34.31.0058]; Russian Academy of Sciences; Russian Federation "Leading Science School" [3951.2012.4]
Место публикации : Anal. Bioanal. Chem.: SPRINGER HEIDELBERG, 2014. - Vol. 406, Is. 23. - С. 5715-5726. - ISSN 1618-2642, DOI 10.1007/s00216-014-7986-2. - ISSN 1618-2650
Примечания : Cited References: 67. - This work was supported by RFBR grant 12-04-00131, by the programs of the Government of the Russian Federation "Measures to Attract Leading Scientists to Russian Educational Institutions" (grant 11.G34.31.0058) and "Molecular and Cellular Biology" of the Russian Academy of Sciences, and the grant from the President of the Russian Federation "Leading Science School" (3951.2012.4).
Предметные рубрики: LIGHT-SENSITIVE PHOTOPROTEIN
CTENOPHORE BEROE ABYSSICOLA
GREEN-FLUORESCENT PROTEIN
INTRACELLULAR CALCIUM
SEQUENCE-ANALYSIS
CA-2+-ACTIVATED PHOTOPROTEIN
CA2+-BINDING PHOTOPROTEIN
SEMISYNTHETIC AEQUORINS
LUMINESCENT PROTEIN
RECOMBINANT OBELIN
Ключевые слова (''Своб.индексиров.''): calcium--coelenterazine--aequorin--obelin--clytin--mitrocomin
Аннотация: Calcium ion is a ubiquitous intracellular messenger, performing this function in many eukaryotic cells. To understand calcium regulation mechanisms and how disturbances of these mechanisms are associated with disease states, it is necessary to measure calcium inside cells. Ca2+-regulated photoproteins have been successfully used for this purpose for many years. Here we report the results of comparative studies on the properties of recombinant aequorin from Aequorea victoria, recombinant obelins from Obelia geniculata and Obelia longissima, recombinant mitrocomin from Mitrocoma cellularia, and recombinant clytin from Clytia gregaria as intracellular calcium indicators in a set of identical in vitro and in vivo experiments. Although photoproteins reveal a high degree of identity of amino acid sequences and spatial structures, and, apparently, have a common mechanism for the bioluminescence reaction, they were found to differ in the Ca2+ concentration detection limit, the sensitivity of bioluminescence to Mg2+, and the rates of the rise of the luminescence signal with a sudden change of Ca2+ concentration. In addition, the bioluminescence activities of Chinese hamster ovary cells expressing wild-type photoproteins also differed. The light signals of cells expressing mitrocomin, for example, slightly exceeded the background, suggesting that mitrocomin may be hardly used to detect intracellular Ca2+ without modifications improving its properties. On the basis of experiments on the activation of endogenous P2Y(2) receptor in Chinese hamster ovary cells by ATP, we suggest that wild-type aequorin and obelin from O. longissima are more suitable for calcium detection in cytoplasm, whereas clytin and obelin from O. geniculata can be used for calcium measurement in cell compartments with high Ca2+ concentration.
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20.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Navdaev A..., Subramanian H..., Petunin A..., Clemetson K.J., Gambaryan S..., Walter U...
Заглавие : Echicetin Coated Polystyrene Beads: A Novel Tool to Investigate GPIb-Specific Platelet Activation and Aggregation
Колич.характеристики :10 с
Коллективы : DFG [SFB688, TP A2, GA 1561/1-1]; BMBF [01EO1003]; Rudolf-Marx-stipendium; Gesellschaft fur Thrombose- und Hamostaseforschung (GTH)
Место публикации : PLoS One: PUBLIC LIBRARY SCIENCE, 2014. - Vol. 9, Is. 4. - Ст.e93569. - ISSN 1932-6203, DOI 10.1371/journal.pone.0093569
Примечания : Cited References: 36. - This study was supported by DFG (SFB688, TP A2, and grant GA 1561/1-1), BMBF (01EO1003) and Rudolf-Marx-stipendium 2014 funded by Gesellschaft fur Thrombose- und Hamostaseforschung (GTH). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Предметные рубрики: VON-WILLEBRAND-FACTOR
GLYCOPROTEIN IB-ALPHA
DEPENDENT PROTEIN-KINASE
SIGNALING PATHWAY
THROMBUS FORMATION
INTEGRIN ALPHA(IIB)BETA(3)
VONWILLEBRAND-FACTOR
HIGH-SHEAR
IGM-KAPPA
RISTOCETIN
Аннотация: von Willebrand factor/ristocetin (vWF/R) induces GPIb-dependent platelet agglutination and activation of alpha IIb beta 3 integrin, which also binds vWF. These conditions make it difficult to investigate GPIb-specific signaling pathways in washed platelets. Here, we investigated the specific mechanisms of GPIb signaling using echicetin-coated polystyrene beads, which specifically activate GPIb. We compared platelet activation induced by echicetin beads to vWF/R. Human platelets were stimulated with polystyrene beads coated with increasing amounts of echicetin and platelet activation by echicetin beads was then investigated to reveal GPIb specific signaling. Echicetin beads induced alpha IIb beta 3-dependent aggregation of washed platelets, while under the same conditions vWF/R treatment led only to alpha IIb beta 3-independent platelet agglutination. The average distance between the echicetin molecules on the polystyrene beads must be less than 7 nm for full platelet activation, while the total amount of echicetin used for activation is not critical. Echicetin beads induced strong phosphorylation of several proteins including p38, ERK and PKB. Synergistic signaling via P2Y(12) and thromboxane receptor through secreted ADP and TxA(2), respectively, were important for echicetin bead triggered platelet activation. Activation of PKG by the NO/sGC/cGMP pathway inhibited echicetin bead-induced platelet aggregation. Echicetin-coated beads are powerful and reliable tools to study signaling in human platelets activated solely via GPIb and GPIb-triggered pathways.
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