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Вид документа : Однотомное издание
Шифр издания : А.с. 1035962!-817652
Автор(ы) : Межевикин В.В., Высоцкий Е.С., Заворуев В.В., Родичева Э.К.
Заглавие : Способ получения препарата бактериальной люциферазы .-
Выходные данные : Б.м.,Б.г.
Коллективы : Ин-т биофиз. СО АН СССР
Цена : Б.ц.
ГРНТИ : 34.27.51
Предметные рубрики: ЛЮЦЕФЕРАЗА
ПОЛУЧЕНИЕ
СПОСОБ
PHOTOBACTERIUM LEIOGNATHI
БАКТЕРИИ
ENZYME ISOLATION
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Вид документа : Статья из сборника (однотомник)
Шифр издания :
Автор(ы) : Петушков В.Н., Шефер Л.П., Родионова Н.С.
Заглавие : Биолюминесцентный анализ НАДН-зависимых дегидрогеназ : научное издание
Место публикации : Получ. и применение биокатализаторов в нар. х-ве и мед. Тез. докл. 5 Всес. симп. по инж. энзимол., Кобулети, май, 1985. Т. 1. - Олайне, 1985. - С. 168
ГРНТИ : 34.15.19
Предметные рубрики: НАДНДЕГИДРОГЕНАЗА
ОПРЕДЕЛЕНИЕ
БИОЛЮМИНЕСЦЕНТНЫЙ МЕТОД
НАДН-ФМНОКСИДОРЕДУКТАЗА
ЛЮЦИФЕРАЗА
БИОЛЮМИНЕСЦЕНТНАЯ СИСТЕМА
ENZYME SYSTEMS
ANALYTICAL USES
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Purtov K. V., Petushkov V. N., Rodionova N. S., Gitelson J. I.
Заглавие : Why does the bioluminescent fungus Armillaria mellea have luminous mycelium but nonluminous fruiting body?
Место публикации : Doklad. Biochem. Biophys.: Maik Nauka Publishing / Springer SBM, 2017. - Vol. 474, Is. 1. - С. 217-219. - ISSN 16076729 (ISSN) , DOI 10.1134/S1607672917030176
Аннотация: By determining the components involved in the bioluminescence process in luminous and nonluminous organs of the honey fungus Armillaria mellea, we have established causes of partial luminescence of this fungus. The complete set of enzymes and substrates required for bioluminescence is formed only in the mycelium and only under the conditions of free oxygen access. Since the synthesis of luciferin precursor (hispidin) and 3-hydroxyhispidin hydroxylase in the fruiting bodies is blocked, the formation of luciferin—the key component of fungal bioluminescent system—was not observed. That is why the fruiting body of Armillaria mellea is nonluminous despite the presence of luciferase, the enzyme that catalyzes the oxidation of luciferin with a photon emission. © 2017, Pleiades Publishing, Ltd.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Lisitsa A. E., Sukovatyi L. A., Kratasyuk V. A., Nemtseva E. V.
Заглавие : Viscous Media Slow Down the Decay of the Key Intermediate in Bacterial Bioluminescent Reaction
Место публикации : Doklad. Biochem. Biophys.: Pleiades Publishing, 2020. - Vol. 492, Is. 1. - С. 162-165. - ISSN 16076729 (ISSN), DOI 10.1134/S1607672920020106
Аннотация: Abstract: The effects of medium viscosity on the decay rate of the 4a-hydroperoxyflavin intermediate of the bioluminescent reaction was investigated. It was found that at low concentrations of glycerol or sucrose (viscosity 1.1–1.3 cP) the decay rate rises, whereas a further increase in viscosity to 6.2 cP leads to a decrease in the decay rate following a power function with an exponent of 0.82–0.84. Using molecular dynamics methods, it was shown that the presence of glycerol and sucrose molecules causes a change in the mobility of the amino acid residues in the active center of luciferase, particularly those responsible for binding of flavin. The results obtained are indicative of two opposite effects of viscous media with glycerol and sucrose: (1) destabilization of 4a-hydroperoxyflavin due to a change in the structural and dynamic properties of the protein and (2) stabilization of this intermediate by the decrease in the diffusion rate of its decay products. © 2020, Pleiades Publishing, Ltd.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Lisitsa A. E., Sukovatyi L. A., Kratasyuk V. A., Nemtseva E. V.
Заглавие : Viscous Media Slow Down the Decay of the Key Intermediate in Bacterial Bioluminescent Reaction
Колич.характеристики :4 с
Коллективы : Ministry of Science and Higher Education of the Russian Federation [6.7734.2017, 01201351504]
Место публикации : Dokl. Biochem. Biophys.: MAIK NAUKA/INTERPERIODICA/SPRINGER, 2020. - Vol. 492, Is. 1. - С. 162-165. - ISSN 1607-6729, DOI 10.1134/S1607672920020106. - ISSN 1608-3091(eISSN)
Примечания : Cited References:15. - This work was supported by the Ministry of Science and Higher Education of the Russian Federation (project nos. 6.7734.2017 and 01201351504).
Предметные рубрики: LUCIFERASE
Аннотация: The effects of medium viscosity on the decay rate of the 4a-hydroperoxyflavin intermediate of the bioluminescent reaction was investigated. It was found that at low concentrations of glycerol or sucrose (viscosity 1.1-1.3 cP) the decay rate rises, whereas a further increase in viscosity to 6.2 cP leads to a decrease in the decay rate following a power function with an exponent of 0.82-0.84. Using molecular dynamics methods, it was shown that the presence of glycerol and sucrose molecules causes a change in the mobility of the amino acid residues in the active center of luciferase, particularly those responsible for binding of flavin. The results obtained are indicative of two opposite effects of viscous media with glycerol and sucrose: (1) destabilization of 4a-hydroperoxyflavin due to a change in the structural and dynamic properties of the protein and (2) stabilization of this intermediate by the decrease in the diffusion rate of its decay products.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Tarasova A..., Kudryasheva N..., Kovel E..., Churilov G..., Vnukova N..., Isakova V..., Osipova I...
Заглавие : Use of bioluminescent enzyme system to detect antioxidant activity of fullerenol C60Oy(OH)(x)
Колич.характеристики :2 с
Место публикации : Luminescence: WILEY-BLACKWELL, 2014. - Vol. 29. - С. 100-101. - ISSN 1522-7235. - ISSN 1522-7243
Примечания : Cited References: 6
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Larionova, Marina D., Markova, Svetlana V., Vysotski, Eugene S.
Заглавие : Tyr72 and Tyr80 are Involved in the Formation of an Active Site of a Luciferase of Copepod Metridia longa
Колич.характеристики :8 с
Коллективы : Russian Science Foundation [14-14-01119]
Место публикации : Photochem. Photobiol.: WILEY, 2017. - Vol. 93, Is. 2. - С. 503-510. - ISSN 0031-8655, DOI 10.1111/php.12694. - ISSN 1751-1097(eISSN)
Примечания : Cited References:41. - This work was supported by the grant 14-14-01119 of the Russian Science Foundation.
Предметные рубрики: CA2+-REGULATED PHOTOPROTEIN OBELIN
COELENTERAZINE-BINDING PROTEIN
Аннотация: Luciferase of copepod Metridia longa (MLuc) is a naturally secreted enzyme catalyzing the oxidative decarboxylation of coelenterazine with the emission of light. To date, three nonallelic isoforms of different lengths (17-24 kDa) for M. longa luciferase have been cloned. All the isoforms are single-chain proteins consisting of a 17-residue signal peptide for secretion, variable N-terminal part and conservative C-terminus responsible for luciferase activity. In contrast to other bioluminescent proteins containing a lot of aromatic residues which are frequently involved in light emission reaction, the C-terminal part of MLuc contains only four Phe, two Tyr, one Trp and two His residues. To figure out whether Tyr residues influence bioluminescence, we constructed the mutants with substitution of Tyr to Phe (Y72F and Y80F). Tyrosine substitutions do not eliminate the ability of luciferase to bioluminescence albeit significantly reduce relative specific activity and change bioluminescence kinetics. In addition, the Tyr replacements have no effect on bioluminescence spectrum, thereby indicating that tyrosines are not involved in the emitter formation. However, as it was found that the intrinsic fluorescence caused by Tyr residues is quenched by a reaction substrate, coelenterazine, in concentration-dependent manner, we infer that both tyrosine residues are located in the luciferase substrate-binding cavity.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Kovel E. S., Kicheeva A. G., Vnukova N. G., Churilov G. N., Stepin E. A., Kudryasheva N. S.
Заглавие : Toxicity and antioxidant activity of fullerenol c60,70 with low number of oxygen substituents
Место публикации : Int. J. Mol. Sci.: MDPI AG, 2021. - Vol. 22, Is. 12. - Ст.6382. - ISSN 16616596 (ISSN), DOI 10.3390/ijms22126382
Аннотация: Fullerene is a nanosized carbon structure with potential drug delivery applications. We studied the bioeffects of a water-soluble fullerene derivative, fullerenol, with 10-12 oxygen groups (F10-12); its structure was characterized by IR and XPS spectroscopy. A bioluminescent enzyme system was used to study toxic and antioxidant effects of F10-12 at the enzymatic level. Antioxidant characteristics of F10-12 were revealed in model solutions of organic and inorganic oxidizers. Low-concentration activation of bioluminescence was validated statistically in oxidizer solutions. Toxic and antioxidant characteristics of F10-12 were compared to those of homologous fullerenols with a higher number of oxygen groups:F24-28 and F40-42. No simple dependency was found between the toxic/antioxidant characteristics and the number of oxygen groups on the fullerene’s carbon cage. Lower toxicity and higher antioxidant activity of F24-28 were identified and presumptively attributed to its higher solubility. An active role of reactive oxygen species (ROS) in the bioeffects of F10-12 was demonstrated. Correlations between toxic/antioxidant characteristics of F10-12 and ROS content were evaluated. Toxic and antioxidant effects were related to the decrease in ROS content in the enzyme solutions. Our results reveal a complexity of ROS effects in the enzymatic assay system. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Kovel, Ekaterina S., Kicheeva, Arina G., Vnukova, Natalia G., Churilov, Grigory N., Stepin, Evsei A., Kudryasheva, Nadezhda S.
Заглавие : Toxicity and Antioxidant Activity of Fullerenol C-60,C-70 with Low Number of Oxygen Substituents
Колич.характеристики :17 с
Коллективы : RFBRRussian Foundation for Basic Research (RFBR) [N18-29-19003]; RFBR, Krasnoyarsk Territory; Krasnoyarsk Regional Fund of Science [N20-44-243001]; Program of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Fundamental Study 2020-2025 (Russian Federation)
Место публикации : Int. J. Mol. Sci.: MDPI, 2021. - Vol. 22, Is. 12. - Ст.6382. - ISSN 1422-0067(eISSN), DOI 10.3390/ijms22126382
Примечания : Cited References:93. - This research was funded by RFBR, N18-29-19003; RFBR, Krasnoyarsk Territory and Krasnoyarsk Regional Fund of Science, N20-44-243001; and partly supported by the Program of the Federal Service for Surveillance on Consumer Rights Protection and HumanWellbeing, Fundamental Study 2020-2025 (Russian Federation).
Предметные рубрики: HUMIC SUBSTANCES
DETOXIFICATION PROCESSES
BIOLOGICAL-ACTIVITY
Аннотация: Fullerene is a nanosized carbon structure with potential drug delivery applications. We studied the bioeffects of a water-soluble fullerene derivative, fullerenol, with 10-12 oxygen groups (F10-12); its structure was characterized by IR and XPS spectroscopy. A bioluminescent enzyme system was used to study toxic and antioxidant effects of F10-12 at the enzymatic level. Antioxidant characteristics of F10-12 were revealed in model solutions of organic and inorganic oxidizers. Low-concentration activation of bioluminescence was validated statistically in oxidizer solutions. Toxic and antioxidant characteristics of F10-12 were compared to those of homologous fullerenols with a higher number of oxygen groups:F24-28 and F40-42. No simple dependency was found between the toxic/antioxidant characteristics and the number of oxygen groups on the fullerene's carbon cage. Lower toxicity and higher antioxidant activity of F24-28 were identified and presumptively attributed to its higher solubility. An active role of reactive oxygen species (ROS) in the bioeffects of F10-12 was demonstrated. Correlations between toxic/antioxidant characteristics of F10-12 and ROS content were evaluated. Toxic and antioxidant effects were related to the decrease in ROS content in the enzyme solutions. Our results reveal a complexity of ROS effects in the enzymatic assay system.
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10.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Shishatskaya E.I., Volova T.G., Puzyr A.P., Mogilnaya O.A., Efremov S.N.
Заглавие : Tissue response to the implantation of biodegradable polyhydroxyalkanoate sutures
Место публикации : Journal of Materials Science: Materials in Medicine. - 2004. - Vol. 15, Is. 6. - С. 719-728. - ISSN 09574530 (ISSN) , DOI 10.1023/B:JMSM.0000030215.49991.0d
Ключевые слова (''Своб.индексиров.''): necrosis--polymeric implants--suppurative inflammation--tissue reaction--biodegradation--calcification (biochemistry)--cells--copolymers--implants (surgical)--silk--tissue--tumors--materials science--acid phosphatase--copolymer--poly(3 hydroxybutyric acid)--polyhydroxyalkanoic acid--polyhydroxyvaleric acid--unclassified drug--animal experiment--animal model--article--biodegradable implant--blood vessel reactivity--catgut--controlled study--enzyme activity--female--giant cell--histochemistry--inflammation--macrophage--nonhuman--priority journal--rat--silk--suture--tensile strength--tissue reaction--tissue structure--wound healing--young modulus--absorbable implants--animals--female--fibrosis--foreign-body reaction--hydroxybutyrates--muscle, skeletal--polyesters--polymers--rats--rats, wistar--sutures--treatment outcome--wounds, penetrating--animalia
Аннотация: Polyhydroxyalkanoate (PHA) sutures were implanted to test animals intramuscularly, and tissue reaction was investigated and compared with the reaction to silk and catgut. Tested monofilament sutures made of PHAs of two types polyhydroxybutyrate (PHB) and a copolymer of hydroxybutyrate and hydroxyvalerate (PHV) featured the strength necessary for the healing of muscle-fascial wounds. The reaction of tissues to polymeric implants was similar to their reaction to silk and was less pronounced than the reaction to catgut; it was expressed in a transient post-traumatic inflammation (up to four weeks) and the formation of a fibrous capsule less than 200 ?m thick, which became as thin as 4060 ?m after 16 weeks, in the course of reverse development. Macrophages and foreign-body giant cells with a high activity of acid phosphatase were actively involved in this process. PHB and PHB/PHV sutures implanted intramuscularly for an extended period (up to one year) did not cause any acute vascular reaction at the site of implantation or any adverse events, such as suppurative inflammation, necrosis, calcification of the fibrous capsule or malignant tumor formation. No statistically significant differences were revealed in the tissue response to polymer sutures of the two types. Capsules around silk and catgut sutures did not become significantly thinner.
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11.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Shishatskaya E.I., Volova T.G., Efremov S.N., Puzyr' A.P., Mogil'Naya O.A.
Заглавие : Tissue response to biodegradable suture threads made of polyhydroxyalkanoates
Место публикации : Biomedical Engineering. - 2002. - Vol. 36, Is. 4. - С. 210-217. - ISSN 00063398 (ISSN) , DOI 10.1023/A:1021184119268
Ключевые слова (''Своб.индексиров.''): acid phosphatase--alkaline phosphatase--polyhydroxyalkanoic acid--animal experiment--animal tissue--article--biocompatibility--biodegradability--controlled study--elasticity--enzyme activity--enzyme mechanism--female--histochemistry--incision--nonhuman--physical chemistry--postoperative period--rat--rigidity--suture--thickness--tissue reaction--wound healing--animalia
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12.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Lonshakova-Mukina, Victoria I., Esimbekova, Elena N., Kratasyuk, Valentina A.
Заглавие : Thermal Inactivation of Butyrylcholinesterase in Starch and Gelatin Gels
Колич.характеристики :10 с
Коллективы : Government of Krasnoyarsk Territory; Krasnoyarsk Regional Fund of Science; Russian Foundation for Basic ResearchRussian Foundation for Basic Research (RFBR) [20-44-242001]
Место публикации : Catalysts: MDPI, 2021. - Vol. 11, Is. 4. - Ст.492. - ISSN 2073-4344(eISSN), DOI 10.3390/catal11040492
Примечания : Cited References:39. - The research was funded by the Government of Krasnoyarsk Territory, Krasnoyarsk Regional Fund of Science, and the Russian Foundation for Basic Research [project No 20-44-242001].
Аннотация: The present study demonstrates a simple approach to enhancing thermal stability of butyrylcholinesterase (BChE) by using natural polymers. Analysis of thermal inactivation of the tetrameric BChE in starch and gelatin gels at 50-64 degrees C showed that thermal inactivation followed second-order kinetics and involved two alternating processes of BChE inactivation, which occurred at different rates (fast and slow processes). The activation enthalpy Delta H-# and the activation entropy Delta S-# for BChE in starch and gelatin gels were evaluated. The values of Delta H-# for the fast and the slow thermal inactivation of BChE in starch gel were 61 +/- 3, and 22 +/- 2 kcal/mol, respectively, and the values of Delta S-# were 136 +/- 12 and -2.03 +/- 0.05 cal center dot K-1 center dot mol(-1), respectively. Likewise, the values of Delta H-# for BChE in gelatin gel were 58 +/- 6 and 109 +/- 11 kcal/mol, and the values of Delta S-# were 149 +/- 16 and 262 +/- 21 cal center dot K-1 center dot mol(-1), respectively. The values of the activation parameters obtained in this study suggest that starch gel produced a stronger stabilizing effect on BChE exposed to elevated temperatures over long periods compared with gelatin gel.
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13.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Kratasyuk V.A., Esimbekova E.N., Gladyshev M.I., Khromichek E.B., Kuznetsov A.M., Ivanova E.A.
Заглавие : The use of bioluminescent biotests for study of natural and laboratory aquatic ecosystems
Место публикации : Chemosphere. - 2001. - Vol. 42, Is. 8. - С. 909-915. - ISSN 00456535 (ISSN) , DOI 10.1016/S0045-6535(00)00177-6
Ключевые слова (''Своб.индексиров.''): alcohol dehydrogenase--bacterial luciferase--bioluminescence--blooming--pollution--trypsin--water toxicity--alcohol dehydrogenase--benzoquinone--luciferase--trypsin--aquatic ecosystem--bioluminescence--water quality--article--bacterium culture--bioluminescence--blue green alga--ecosystem--pond--seasonal variation--water pollution--water quality--benzoquinones--biological assay--cyanobacteria--ecosystem--environmental monitoring--eutrophication--fmn reductase--indicators and reagents--luminescent measurements--nadh, nadph oxidoreductases--water pollutants--russian federation--algae--bacteria (microorganisms)--chlorophyta--cyanobacteria--uncultured cyanobacterium
Аннотация: A set of bioluminescent tests was developed to monitor water quality in natural and laboratory ecosystems. It consisted of four bioluminescent systems: luminous bacteria, coupled enzyme system NADH:FMN-oxidoreductase-luciferase and triplet enzyme systems with alcohol dehydrogenase and trypsin. The set of biotests was applied for a small forest pond (Siberia, Russia), laboratory microecosystems polluted with benzoquinone and a batch culture of blue-green algae. Thereby effects of natural water compared to those of models of heavy pollution and "bloom" of blue-greens on the bioluminescent tests were revealed. The set of biotests was not affected by a natural seasonal variability of water quality in the unpolluted pond, but responded to the heavy pollution and the "bloom" of blue-greens. The set of biotests could be recommended as the alarm test to control the acute toxicity of natural water bodies. В© 2001 Elsevier Science Ltd.
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14.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Larionova, Marina D., Markova, Svetlana, V, Tikunova, Nina, V, Vysotski, Eugene S.
Заглавие : The Smallest Isoform ofMetridia longaLuciferase as a Fusion Partner for Hybrid Proteins
Колич.характеристики :16 с
Коллективы : Russian Foundation for Basic ResearchRussian Foundation for Basic Research (RFBR) [18-44-242001]; Krasnoyarsk Regional Fund of Science; ICBFM SB RAS [AAAA-A17-117020210027-9]; Government of Krasnoyarsk Territory
Место публикации : Int. J. Mol. Sci.: MDPI, 2020. - Vol. 21, Is. 14. - Ст.4971. - ISSN 1422-0067(eISSN), DOI 10.3390/ijms21144971
Примечания : Cited References:49. - The reported study was funded by the Russian Foundation for Basic Research (No. 18-44-242001), Government of Krasnoyarsk Territory, Krasnoyarsk Regional Fund of Science (S.V.M, M.D.L., and E.S.V.) and by the Russian State funded budget project of ICBFM SB RAS No. AAAA-A17-117020210027-9 (N.V.T.).
Предметные рубрики: COELENTERAZINE-BINDING PROTEIN
BIOLUMINESCENT REPORTER
GAUSSIA
Аннотация: Bioluminescent proteins are widely used as reporter molecules in various in vitro and in vivo assays. The smallest isoform of Metridia luciferase (MLuc7) is a highly active, naturally secreted enzyme which, along with other luciferase isoforms, is responsible for the bright bioluminescence of marine copepodMetridia longa. In this study, we report the construction of two variants of a hybrid protein consisting of MLuc7 and 14D5a single-chain antibody to the surface glycoprotein E of tick-borne encephalitis virus as a model fusion partner. We demonstrate that, whereas fusion of a single-chain antibody to either N- or C-terminus of MLuc7 does not affect its bioluminescence properties, the binding site on the single-chain antibody influences its binding capacity. The affinity of 14D5a-MLuc7 hybrid protein (K-D= 36.2 nM) where the C-terminus of the single-chain antibody was fused to the N-terminus of MLuc7, appeared to be 2.5-fold higher than that of the reverse, MLuc7-14D5a (K-D= 87.6 nM). The detection limit of 14D5a-MLuc7 hybrid protein was estimated to be 45 pg of the recombinant glycoprotein E. Although the smallest isoform ofM. longaluciferase was tested as a fusion partner only with a single-chain antibody, it is reasonable to suppose that MLuc7 can also be successfully used as a partner for genetic fusion with other proteins.
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15.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Larionova M. D., Markova S. V., Tikunova N. V., Vysotski E. S.
Заглавие : The smallest isoform of Metridia longa luciferase as a fusion partner for hybrid proteins
Место публикации : Int. J. Mol. Sci.: MDPI AG, 2020. - Vol. 21, Is. 14. - Ст.4971. - С. 1-16. - ISSN 16616596 (ISSN), DOI 10.3390/ijms21144971
Аннотация: Bioluminescent proteins are widely used as reporter molecules in various in vitro and in vivo assays. The smallest isoform of Metridia luciferase (MLuc7) is a highly active, naturally secreted enzyme which, along with other luciferase isoforms, is responsible for the bright bioluminescence of marine copepod Metridia longa. In this study, we report the construction of two variants of a hybrid protein consisting of MLuc7 and 14D5a single-chain antibody to the surface glycoprotein E of tick-borne encephalitis virus as a model fusion partner. We demonstrate that, whereas fusion of a single-chain antibody to either N-or C-terminus of MLuc7 does not affect its bioluminescence properties, the binding site on the single-chain antibody influences its binding capacity. The affinity of 14D5a-MLuc7 hybrid protein (KD = 36.2 nM) where the C-terminus of the single-chain antibody was fused to the N-terminus of MLuc7, appeared to be 2.5-fold higher than that of the reverse, MLuc7-14D5a (KD = 87.6 nM). The detection limit of 14D5a-MLuc7 hybrid protein was estimated to be 45 pg of the recombinant glycoprotein E. Although the smallest isoform of M. longa luciferase was tested as a fusion partner only with a single-chain antibody, it is reasonable to suppose that MLuc7 can also be successfully used as a partner for genetic fusion with other proteins. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.
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16.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Larionova, Marina D., Markova, Svetlana V., Vysotski, Eugene S.
Заглавие : The novel extremely psychrophilic luciferase from Metridia longa: Properties of a high-purity protein produced in insect cells
Колич.характеристики :7 с
Коллективы : Bayer AG (Germany); Russian Academy of Sciences [01201351504]; RFBR; Government of Krasnoyarsk Territory [16-44-242099]
Место публикации : Biochem. Biophys. Res. Commun.: ACADEMIC PRESS INC ELSEVIER SCIENCE, 2017. - Vol. 483, Is. 1. - С. 772-778. - ISSN 0006-291X, DOI 10.1016/j.bbrc.2016.12.067. - ISSN 1090-2104(eISSN)
Примечания : Cited References:24. - The cloning of cDNAs encoding MLuc2 isoforms of M. longa was supported by Bayer AG (Germany) and the state budget allocated to the fundamental research at the Russian Academy of Sciences (project No. 01201351504); all other studies were funded by RFBR and Government of Krasnoyarsk Territory according to the research project No. 16-44-242099.
Предметные рубрики: CDNA CLONING
EXPRESSION
ENZYME
Ключевые слова (''Своб.индексиров.''): bioluminescence--coelenterazine--bioluminescent reporter--psychrophilic--enzyme--molecular adaptation
Аннотация: The bright bioluminescence of copepod Metridia longa is conditioned by a small secreted coelenterazinedependent luciferase (MLuc). To date, three isoforms of MLuc differing in length, sequences, and some properties were cloned and successfully applied as high sensitive bioluminescent reporters. In this work, we report cloning of a novel group of genes from M. longa encoding extremely psychrophilic isoforms of MLuc (MLuc2-type). The novel isoforms share only similar to 54-64% of protein sequence identity with the previously cloned isoforms and, consequently, are the product of a separate group of paralogous genes. The MLuc2 isoform with consensus sequence was produced as a natively folded protein using baculovirus/ insect cell expression system, purified, and characterized. The MLuc2 displays a very high bioluminescent activity and high thermostability similar to those of the previously characterized M. longa luciferase isoform MLuc7. However, in contrast to MLuc7 revealing the highest activity at 12-17 degrees C and 0.5 M NaCl, the bioluminescence optima of MLuc2 isoforms are at similar to 5 degrees C and 1 M NaCl. The MLuc(2) adaptation to cold is also accompanied by decrease of melting temperature and affinity to substrate suggesting a more conformational flexibility of a protein structure. The luciferase isoforms with different temperature optima may provide adaptability of the M. longa bioluminescence to the changes of water temperature during diurnal vertical migrations. (C) 2016 Elsevier Inc. All rights reserved.
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17.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Kudryasheva N.S., Kudinova I.Y., Esimbekova E.N., Kratasyuk V.A., Stom D.I.
Заглавие : The influence of quinones and phenols on the triple NAD(H)-dependent enzyme systems
Колич.характеристики :8 с
Место публикации : Chemosphere: PERGAMON-ELSEVIER SCIENCE LTD, 1999. - Vol. 38, Is. 4. - P751-758. - ISSN 0045-6535, DOI 10.1016/S0045-6535(98)00218-5
Примечания : Cited References: 7
Аннотация: Kinetics of the triple bioluminescent enzyme system: alcohol dehydrogenase - NADH:FMN-oxidoreductase - luciferase in the presence of quinones and phenols has been studied. The correspondence between the bioluminescent kinetic parameters, redox potentials and concentrations of the quinones and phenols has been estimated. The substances have been shown to change bioluminescent kinetics through moving off the NAD(+)/NADH balance in the enzyme processes. This system is proposed to be used as enzymatic biotest in ecological monitoring. (C) 1998 Elsevier Science Ltd. All rights reserved.
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18.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Sukovatyi L. A., Lisitsa A. E., Kratasyuk V. A., Nemtseva E. V.
Заглавие : The Effect of Osmolytes on the Bioluminescent Reaction of Bacteria: Structural and Dynamic Properties
Место публикации : Biophysics: Pleiades journals, 2020. - Vol. 65, Is. 6. - С. 966-971. - ISSN 00063509 (ISSN), DOI 10.1134/S0006350920060202
Аннотация: The effects of viscous media with glycerol and sucrose (10–40%) on the kinetics of the bacterial bioluminescent reaction have been investigated by stopped-flow technique. Increment of quantum yield in media with 10% of both osmolytes was shown. Higher concentrations of glycerol, up to 30–40%, were found to reduce the efficiency of the reaction, while this effect was not observed in the media with sucrose. The molecular dynamics simulation was used to study the structure of bacterial luciferase surrounded by either water molecules solely or by mixture of water with various numbers of glycerol/sucrose molecules. It was found that both cosolvents at studied concentrations did not cause a significant change in conformation of bacterial luciferase. The calculated root-mean-square fluctuation for C?-atoms of bacterial luciferase ?-subunit indicated that the higher flexibility of the enzyme mobile loop could be responsible for increment of quantum yield in the presence of 10% of both osmolytes. The active site of bacterial luciferase was found to be accessible for glycerol molecules while sucrose did not enter catalytic gorge. Moreover, at 30 and 40% concentration the glycerol molecules were found to locate in the active site of bacterial luciferase throughout the whole simulation time. © 2020, Pleiades Publishing, Inc.
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19.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Markova, Svetlana V., Larionova, Marina D., Gorbunova, Darya A., Vysotski, Eugene S.
Заглавие : The disulfide-rich Metridia luciferase refolded from E. coli inclusion bodies reveals the properties of a native folded enzyme produced in insect cells
Колич.характеристики :7 с
Коллективы : RFBR; Government of Krasnoyarsk Territory [16-44-242099]; Russian Academy of Sciences [0356-2016-0712]
Место публикации : J. Photochem. Photobiol. B-Biol.: ELSEVIER SCIENCE SA, 2017. - Vol. 175. - С. 51-57. - ISSN 1011-1344, DOI 10.1016/j.jphotobiol.2017.08.024
Примечания : Cited References:30. - These studies were funded by RFBR and the Government of Krasnoyarsk Territory according to the research project No. 16-44-242099 and the state budget allocated to the fundamental research at the Russian Academy of Sciences (project No. 0356-2016-0712).
Предметные рубрики: GAUSSIA-PRINCEPS LUCIFERASE
ESCHERICHIA-COLI
EXPRESSION
PROTEIN
Ключевые слова (''Своб.индексиров.''): copepod luciferase--disulfide bonds--cysteine-rich protein--oxidative--refolding
Аннотация: The bioluminescence of a marine copepod Metridia Tonga is determined by a small secreted coelenterazine-dependent luciferase that uses coelenterazine as a substrate of enzymatic reaction to generate light (lambda(max) = 480 nm). To date, four different isoforms of the luciferase differing in size, sequences, and properties have been cloned by functional screening. All of them contain ten conserved Cys residues that suggests up to five S-S intramolecular bonds per luciferase molecule. Whereas the use of copepod luciferases as bioluminescent reporters in biomedical research in vivo is growing from year to year, their application for in vitro assays is still limited by the difficulty in obtaining significant amounts of luciferase. The most cost-effective host for producing recombinant proteins is Escherichia coli. However, prokaryotic and eukaryotic cells maintain the reductive environment in cytoplasm that hinders the disulfide bond formation and consequently the proper folding of luciferase. Here we report the expression of the MLuc7 isoform of M. longa luciferase in E. colt cells and the efficient procedure for refolding from inclusion bodies yielding a high-active monomeric protein. Furthermore, in a set of identical experiments we demonstrate that bioluminescent and structural features of MLuc7 produced in bacterial cells are identical to those of MLuc7 isoform produced from culture medium of insect cells. Although the yield of high-purity protein is only 6 mg/L, the application of E. coil cells to produce the luciferase is simpler and more cost-effective than the use of insect cells. We expect that the suggested technology of Metridia luciferase production allows obtaining of sufficient amounts of protein both for the development of novel in vitro analytical assays with the use of MLuc7 as a label and for structural studies.
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20.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Chang D., Liu Y., Chen Y., Hu X., Burov A., Puzyr A., Bondar V., Yao L.
Заглавие : Study of the immunogenicity of the VP2 protein of canine parvovirus produced using an improved Baculovirus expression system
Место публикации : BMC Vet. Res.: BioMed Central Ltd., 2020. - Vol. 16, Is. 1. - Ст.202. - ISSN 17466148 (ISSN), DOI 10.1186/s12917-020-02422-3
Аннотация: Background: Canine parvovirus (CPV) is now recognized as a serious threat to the dog breeding industry worldwide. Currently used CPV vaccines all have their specific drawbacks, prompting a search for alternative safe and effective vaccination strategies such as subunit vaccine. VP2 protein is the major antigen targeted for developing CPV subunit vaccine, however, its production in baculovirus expression system remains challenging due to the insufficient yield. Therefore, our study aims to increase the VP2 protein production by using an improved baculovirus expression system and to evaluate the immunogenicity of the purified VP2 protein in mice. Results: The results showed that high-level expression of the full length VP2 protein was achieved using our modified baculovirus expression system. The recombinant virus carrying two copies of VP2 gene showed the highest expression level, with a productivity of 186 mg/L, which is about 1.4-1.6 fold that of the recombinant viruses carrying only one copy. The purified protein reacted with Mouse anti-His tag monoclonal antibody and Rabbit anti-VP2 polyclonal antibody. BALB/c mice were intramuscularly immunized with purified VP2 protein twice at 2 week intervals. After vaccination, VP2 protein could induce the mice produce high level of hemagglutination inhibition antibodies. Conclusions: Full length CPV VP2 protein was expressed at high level and purified efficiently. Moreover, it stimulated mice to produce high level of antibodies with hemmaglutination inhibition properties. The VP2 protein expressed in this study could be used as a putative economic and efficient subunit vaccine against CPV infection. © 2020 The Author(s).
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