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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Frank L.A., Borisova V.V., Markova S.V., Malikova N.P., Stepanyuk G.A., Vysotski E.S.
Заглавие : Violet and greenish photoprotein obelin mutants for reporter applications in dual-color assay
Колич.характеристики :6 с
Место публикации : Anal. Bioanal. Chem.: SPRINGER HEIDELBERG, 2008. - Vol. 391, Is. 8. - С. 2891-2896. - ISSN 1618-2642, DOI 10.1007/s00216-008-2223-5
Примечания : Cited References: 22
Предметные рубрики: ANGSTROM RESOLUTION
RECOMBINANT OBELIN
CRYSTAL-STRUCTURE
BIOLUMINESCENCE
AEQUORIN
IMMUNOASSAY
EXPRESSION
CDNA
PURIFICATION
CLONING
Ключевые слова (''Своб.индексиров.''): ca(2+)-regulated photoprotein--bioluminescence--dual-color assay
Аннотация: Two kinds of Ca(2+)-regulated photoprotein obelin with altered color of bioluminescence were obtained by active-center amino acid substitution. The mutant W92F-H22E emits violet light (lambda(max)=390 nm) and the mutant Y139F emits greenish light (lambda (max)=498 nm), with small spectral overlap, both display high activity and stability and thus may be used as reporters. For demonstration, the mutants were applied in dual-color simultaneous immunoassay of two gonadotropic hormones-follicle-stimulating hormone and luteinizing hormone. Bioluminescence of the reporters was simultaneously triggered by single injection of Ca(2+) solution, divided using band-pass optical filters and measured with a two-channel photometer. The sensitivity of simultaneous bioluminescence assay was close to that of a separate radioimmunoassay.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Puzyr A. P., Burov A. E., Medvedeva S. E., Burova O. G., Bondar V. S.
Заглавие : Two forms of substrate for the bioluminescent reaction in three species of basidiomycetes
Место публикации : Mycol.: Taylor and Francis Ltd., 2019. - Vol. 10, Is. 2. - С. 84-91. - ISSN 21501203 (ISSN) , DOI 10.1080/21501203.2019.1583688
Ключевые слова (''Своб.индексиров.''): cold and hot extracts--culture liquid--enzymatic system--hispidin--luminous fungi--substrate of luminescent reaction
Аннотация: The luminescent response of the enzymatic system of Armillaria borealis on the cold and hot extracts from cell-free culture liquids of Inonotus obliquus, Pholiota sp. and A. borealis was examined. The greatest influence on the light emission produced by the luminescent system of A. borealis was provided by the temperature at which the probes were prepared for assay. Boiling a culture liquid on water bath for a few minutes promoted a multifold increase in the luminescence. The results of luminescence assay suggest that the substance involved in the bioluminescent reaction in higher fungi is presented in culture liquids and mycelia in two forms. In one form, it is ready to interact with the enzymatic system and in the second form, it becomes accessible for the reaction after heat treatment. The pool of thermoactivated substance was found to be much large than the amount of the ready accessible one. We suggest that predecessors of hispidin, which is fungal luciferin precursor, are responsible for this phenomenon. They are not involved in bioluminescence at their original state and are converted into the substrate under the influence of high temperature. © 2019, © 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
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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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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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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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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Krasitskaya V. V., Bashmakova E. E., Kudryavtsev A. N., Vorobjeva M. A., Shatunova E. A., Frank L. A.
Заглавие : The Hybrid Protein ZZ-OL as an Analytical Tool for Biotechnology Research
Колич.характеристики :7 с
Коллективы : Russian FederationRussian Federation [MK-772.2020.4]; Russian Science FoundationRussian Science Foundation (RSF) [16-14-10296]
Место публикации : Russ. J. Bioorg. Chem.: MAIK NAUKA/INTERPERIODICA/SPRINGER, 2020. - Vol. 46, Is. 6. - С. 1004-1010. - ISSN 1068-1620, DOI 10.1134/S106816202006014X. - ISSN 1608-330X(eISSN)
Примечания : Cited References:14. - The study was partially supported by a grant of the President of the Russian Federation for Young Scientists, the Candidates of Sciences (project MK-772.2020.4 in the part involving the synthesis and analysis of variants of proteins with melanoma-inhibiting activity) and a grant of the Russian Science Foundation (project no. 16-14-10296 in the part involving the bioluminescence analysis of binding of DNA aptamers to targets).
Аннотация: The gene of the hybrid protein that encodes the double synthetic fragment proZZ of the immunoglobulin-binding domain of protein A of Staphylococcus aureus and apo-obelin joined by a short linker has been cloned. The corresponding hybrid protein has been obtained by expression in Escherichia coli cells. The protein activated with a substrate (coelenterazine) possesses the bioluminescent Ca2+-dependent activity of the photoprotein close to that of recombinant wild-type obelin, and the immunoglobulin-binding ability of protein A. It has been shown that the hybrid can be used as a highly sensitive label to detect antibodies and estimate their affinity and interaction with recombinant proteins, as well as in investigations of other kinds.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Krasitskaya, Vasilisa V., Goncharova, Natalia S., Biriukov, Vladislav V., Bashmakova, Eugenia E., Kabilov, Marsel R., Baykov, Ivan K., Sokolov, Aleksey E., Frank, Ludmila A.
Заглавие : The Ca2+-Regulated Photoprotein Obelin as a Tool for SELEX Monitoring and DNA Aptamer Affinity Evaluation
Колич.характеристики :6 с
Коллективы : Russian Foundation for Basic Research (RFBR)Russian Foundation for Basic Research (RFBR) [18-38-00531]
Место публикации : Photochem. Photobiol.: WILEY, 2020. - Article in press. - ISSN 0031-8655, DOI 10.1111/php.13274. - ISSN 1751-1097(eISSN)
Примечания : Cited References:25. - This work has been supported by the Russian Foundation for Basic Research (RFBR) under the grant no 18-38-00531.
Предметные рубрики: CARDIAC TROPONIN-I
BIOLUMINESCENCE
IMMUNOASSAY
APTASENSOR
DIAGNOSIS
Аннотация: Bioluminescent solid-phase analysis was proposed to monitor the selection process and to determine binding characteristics of the aptamer-target complexes during design and development of the specific aptamers. The assay involves Ca2+-regulated photoprotein obelin as a simple, sensitive and fast reporter. Applicability and the prospects of the approach were exemplified by identification of DNA aptamers to cardiac troponin I, a highly specific early biomarker for acute myocardial infarction. Two structurally different aptamers specific to various epitopes of troponin I were obtained and then tested in a model bioluminescent assay.
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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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10.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Bashmakova E. E., Krasitskaya V. V., Frank L. A.
Заглавие : Simultaneous genotyping of four single nucleotide polymorphisms associated with risk factors of hemostasis disorders
Место публикации : Comb. Chem. High Throughput Screen. - 2015. - Vol. 18, Is. 10. - С. 930-936. - ISSN 13862073 (ISSN)
Ключевые слова (''Своб.индексиров.''): bioluminescent microassay--multiplex pcr--pext reaction--photoprotein obelin--snp detection
Аннотация: Multiplex simultaneous genotyping technique was developed for four polymorphisms in genes coding for blood coagulation factors and homocysteine metabolism which are considered as thrombophilia related mutations: FV Leiden, FII G20210A, MTHFR C677T, and FVII G10976A. It is based on primer extension reaction with the following bioluminescent solid-phase microassay. At that, two different in bioluminescence obelin mutants were applied to simultaneous detection of two gene allelic variants. The assay is carried out in microtiter plate format and provides fast and reliable genotyping of four single nucleotide polymorphisms in four different genes within 2.5 hours. A large number of clinical samples were analyzed and the obtained results were found to be in complete correlation with those obtained by using conventional RT-PCR techniques. © 2015 Bentham Science Publishers.
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11.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Bashmakova, Eugenia E., Krasitskaya, Vasilisa V., Frank, Ludmila A.
Заглавие : Simultaneous Genotyping of Four Single Nucleotide Polymorphisms Associated with Risk Factors of Hemostasis Disorders
Колич.характеристики :7 с
Коллективы : Russian Science Foundation [14-14-01119]
Место публикации : Comb. Chem. High Throughput Screen: BENTHAM SCIENCE PUBL LTD, 2015. - Vol. 18, Is. 10. - С. 930-936. - ISSN 1386-2073, DOI 10.2174/1386207318666150917095903. - ISSN 1875-5402(eISSN)
Примечания : Cited References:20. - The study was supported by the grant 14-14-01119 of the Russian Science Foundation.
Предметные рубрики: ALLELE-SPECIFIC PCR
FACTOR-V-LEIDEN
BIOLUMINESCENT IMMUNOASSAY
Ключевые слова (''Своб.индексиров.''): snp detection--pext reaction--photoprotein obelin--bioluminescent--microassay--multiplex pcr
Аннотация: Multiplex simultaneous genotyping technique was developed for four polymorphisms in genes coding for blood coagulation factors and homocysteine metabolism which are considered as thrombophilia related mutations: FV Leiden, FII G20210A, MTHFR C677T, and FVII G10976A. It is based on primer extension reaction with the following bioluminescent solid-phase microassay. At that, two different in bioluminescence obelin mutants were applied to simultaneous detection of two gene allelic variants. The assay is carried out in microtiter plate format and provides fast and reliable genotyping of four single nucleotide polymorphisms in four different genes within 2.5 hours. A large number of clinical samples were analyzed and the obtained results were found to be in complete correlation with those obtained by using conventional RT-PCR techniques.
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12.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Kirillova M. A., Ranjan R., Esimbekova E. N., Kratasyuk V. A.
Заглавие : Role of Hsp90 and ATP in modulating apyrase activity and firefly luciferase kinetics
Место публикации : Int. J. Biol. Macromol.: Elsevier B.V., 2019. - Vol. 131. - С. 691-696. - ISSN 01418130 (ISSN) , DOI 10.1016/j.ijbiomac.2019.03.110
Ключевые слова (''Своб.индексиров.''): bioluminescence--heat shock protein 90--high-throughput screening--adenosine triphosphate--apyrase--bovine serum albumin--firefly luciferase--heat shock protein 90--stabilizing agent--article--bioluminescence--clinical study--conformation--controlled study--denaturation--enzyme activity--enzyme kinetics--high throughput screening--incubation time--nonhuman--protein protein interaction--protein refolding--temperature--thermal denaturation--time
Аннотация: The present manuscript describes a novel bioassay consisting of apyrase and heat shock protein 90 (Hsp90) without additional co-chaperone supplementation; intended for high-throughput screening of anti-cancer drugs and prognosis of stress. In this regard, Hsp90 and adenosine 5?-triphosphate (ATP) mediated firefly luciferase (FLuc) kinetics was investigated using apyrase and FLuc as client proteins. Bioluminescent assay containing Hsp90, ATP, and apyrase led to complete loss of luminescence at 50 °C which indicates the protective role of Hsp90 against thermal denaturation. Similarly, the assay sample comprising Hsp90, ATP, and FLuc showed 2 fold increments in luminescence than their counterparts. Introduction of bovine serum albumin (BSA) to the pre-incubated assay mixture led to an initial rise in the luminescence (28%) in comparison to the sample containing Hsp90, ATP and FLuc. Therefore, FLuc based HTS assays are not suitable for clinical samples which may contain stabilizing agents. However, thermally denatured FLuc and apyrase could not regain their active conformation even when Hsp90 and ATP were introduced in the assay system. This observation justifies the role of Hsp90 to be protective rather than a reparation agent when acts without co-chaperones. © 2019
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13.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Vorobjeva M.A., Krasitskaya V.V., Fokina A.A., Timoshenko V.V., Nevinsky G.A., Venyaminova A.G., Frank L.A.
Заглавие : RNA aptamer against autoantibodies associated with multiple sclerosis and bioluminescent detection probe on its basis
Место публикации : Anal. Chem. - 2014. - Vol. 86, Is. 5. - С. 2590-2594. - ISSN 00032700 (ISSN) , DOI 10.1021/ac4037894
Ключевые слова (''Своб.индексиров.''): autoantibodies--detection probes--developed model--high affinity--laboratory test--microplate assay--multiple sclerosis--myelin basic protein--antibodies--rna--bioluminescence
Аннотация: Nowadays, there are no specific laboratory tests for establishing the diagnosis of multiple sclerosis (MS). The presence of proteolytic autoantibodies against myelin basic protein is now considered as a characteristic feature of MS. New 2?-F-containing RNA aptamer of high affinity and specificity to these antibodies was selected. Covalent conjugate of this aptamer and Ca 2+-regulated photoprotein obelin was obtained for the first time and applied as a label in bioluminescent microplate assay to detect target antibodies. The developed model solid-phase microassay is simple, fast, and highly sensitive. © 2014 American Chemical Society.
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14.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Bondar V.S., Ronzhin N.O., Mamaeva E.S., Baron A.V., Gitelson J.I.
Заглавие : Reusable biochemical diagnosis systems based on nanodiamonds
Место публикации : Doklady Biochemistry and Biophysics. - 2013. - Vol. 448, Is. 1. - С. 55-58. - ISSN 16076729 (ISSN) , DOI 10.1134/S160767291301016X
Ключевые слова (''Своб.индексиров.''): cholesterol--diamond--nanoparticle--article--bioassay--blood--chemical model--chemistry--glucose blood level--human--methodology--biological assay--blood glucose--cholesterol--diamond--humans--models, chemical--nanoparticles
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15.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Davydova, Anna, Krasitskaya, Vasilisa, Vorobjev, Pavel, Timoshenko, Valentina, Tupikin, Alexey, Kabilov, Marsel, Frank, Ludmila, Venyaminova, Alya, Vorobyeva, Mariya
Заглавие : Reporter-recruiting bifunctional aptasensor for bioluminescent analytical assays
Колич.характеристики :7 с
Коллективы : Russian Science FoundationRussian Science Foundation (RSF) [16-14-10296]; Russian State funded budget projects [AAAA-A17-117020210021-7, AAAA-A19-119031890015-0]
Место публикации : RSC Adv.: ROYAL SOC CHEMISTRY, 2020. - Vol. 10, Is. 54. - С. 32393-32399. - ISSN 2046-2069(eISSN), DOI 10.1039/d0ra05117a
Примечания : Cited References:33. - The work was supported by the Russian Science Foundation (grant #16-14-10296), Russian State funded budget projects #AAAA-A17-117020210021-7 to ICBFM SB RAS and #AAAA-A19-119031890015-0 to IBP SB RAS.
Предметные рубрики: DNA APTAMER
RNA APTAMER
OBELIN
PURIFICATION
EXPRESSION
SEQUENCES
Аннотация: We report a novel bioluminescent aptasensor, which consists of 2 '-F-RNA aptamer modules joined into a bi-specific aptamer construct. One aptamer module binds the analyte, then after structural rearrangement the second module recruits non-covalently Ca2+-dependent photoprotein obelin from the solution, thus providing a bioluminescent signal. This concept allows using free protein as a reporter, which brings such advantages as no need for aptamer-protein conjugation, a possibility of thermal re-folding of aptamer component with no harm to a protein, and simpler detection protocol. We developed the new 2 '-F-RNA aptamer for obelin, and proposed the strategy for engineering structure-switching bi-modular aptamer constructs which bind the analyte and the obelin in a sequential manner. With the use of hemoglobin as a model analyte, we showed the feasibility of utilizing the aptasensor in a fast and straightforward bioluminescent microplate assay. With a proper design of a secondary structure, this strategy of aptasensor engineering might be further extended to bi-specific aptamer-based bioluminescent sensors for other analytes of interest.
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16.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Sid'ko A.F.
Заглавие : Remote assay for chlorophyll photosynthetic potential of crops on the example of wheat
Место публикации : Biol. Bull: MAIK NAUKA/INTERPERIODICA, 2004. - Vol. 31, Is. 5. - С. 450-456. - 7. - ISSN 1062-3590, DOI 10.1023/B:BIBU.0000043769.80086.02
Примечания : Cited References: 20
Аннотация: An optical remote assay for biological production of crops in the field during the vegetation period is proposed. Our calculations demonstrate a good correlation between the S(t) value and crop yield (in the range from 0.85 to 0.90); the higher is S(t) for the vegetation period, the higher is the crop yield. S(t) value can be used to calculate the increase in dry and wet biomass for various crops during the whole vegetation period with an error of less than 10-12%. The absolute error of crop yield for 110 cultivars of wheat, oats, and barley during the experimental period was +/-3-3.5 quintal/ha with the mean crop yield of 14-38 quintal/ha.
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17.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Sid'ko A.F.
Заглавие : Remote assay for chlorophyll photosynthetic potential of crops on the example of wheat
Колич.характеристики :7 с
Место публикации : Biol. Bull: MAIK NAUKA/INTERPERIODICA, 2004. - Vol. 31, Is. 5. - P450-456. - ISSN 1062-3590, DOI 10.1023/B:BIBU.0000043769.80086.02
Примечания : Cited References: 20
Аннотация: An optical remote assay for biological production of crops in the field during the vegetation period is proposed. Our calculations demonstrate a good correlation between the S(t) value and crop yield (in the range from 0.85 to 0.90); the higher is S(t) for the vegetation period, the higher is the crop yield. S(t) value can be used to calculate the increase in dry and wet biomass for various crops during the whole vegetation period with an error of less than 10-12%. The absolute error of crop yield for 110 cultivars of wheat, oats, and barley during the experimental period was +/-3-3.5 quintal/ha with the mean crop yield of 14-38 quintal/ha.
WOS
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18.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Bakayan A., Picaud S., Malikova N. P., Tricoire L., Lambolez B., Vysotski E. S., Peyrieras N.
Заглавие : Redquorinxs mutants with enhanced calcium sensitivity and bioluminescence output efficiently report cellular and neuronal network activities
Место публикации : Int. J. Mol. Sci.: MDPI AG, 2020. - Vol. 21, Is. 21. - Ст.7846. - С. 1-22. - ISSN 16616596 (ISSN), DOI 10.3390/ijms21217846
Аннотация: Considerable efforts have been focused on shifting the wavelength of aequorin Ca2+? dependent blue bioluminescence through fusion with fluorescent proteins. This approach has notably yielded the widely used GFP?aequorin (GA) Ca2+ sensor emitting green light, and tdTomato-aequorin (Redquorin), whose bioluminescence is completely shifted to red, but whose Ca2+ sensitivity is low. In the present study, the screening of aequorin mutants generated at twenty?four amino acid positions in and around EF?hand Ca2+?binding domains resulted in the isolation of six aequorin single or double mutants (AequorinXS) in EF2, EF3, and C?terminal tail, which exhibited markedly higher Ca2+ sensitivity than wild?type aequorin in vitro. The corresponding Redquorin mutants all showed higher Ca2+ sensitivity than wild?type Redquorin, and four of them (RedquorinXS) matched the Ca2+ sensitivity of GA in vitro. RedquorinXS mutants exhibited unaltered thermostability and peak emission wavelengths. Upon stable expression in mammalian cell line, all RedquorinXS mutants reported the activation of the P2Y2 receptor by ATP with higher sensitivity and assay robustness than wt?Redquorin, and one, RedquorinXS?Q159T, outperformed GA. Finally, wide?field bioluminescence imaging in mouse neocortical slices showed that RedquorinXS?Q159T and GA similarly reported neuronal network activities elicited by the removal of extracellular Mg2+. Our results indicate that RedquorinXS?Q159T is a red light?emitting Ca2+ sensor suitable for the monitoring of intracellular signaling in a variety of applications in cells and tissues, and is a promising candidate for the transcranial monitoring of brain activities in living mice. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.
Scopus
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19.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Bakayan, Adil, Picaud, Sandrine, Malikova, Natalia P., Tricoire, Ludovic, Lambolez, Bertrand, Vysotski, Eugene S., Peyrieras, Nadine
Заглавие : RedquorinXS Mutants with Enhanced Calcium Sensitivity and Bioluminescence Output Efficiently Report Cellular and Neuronal Network Activities
Колич.характеристики :20 с
Коллективы : Centre National de la Recherche ScientifiqueCentre National de la Recherche Scientifique (CNRS); Agence Nationale de la RechercheFrench National Research Agency (ANR) [ANR-10-INBS-04, ANR-11-EQPX-029]; Fondation pour la Recherche sur le Cerveau/Rotary Club de France; RFBRRussian Foundation for Basic Research (RFBR) [20-04-00085]
Место публикации : Int. J. Mol. Sci.: MDPI, 2020. - Vol. 21, Is. 21. - Ст.7846. - ISSN 1422-0067(eISSN), DOI 10.3390/ijms21217846
Примечания : Cited References:53. - This work was supported by grants from Centre National de la Recherche Scientifique (AAP Prematuration CNRS 2016, to A.B. and N.P.; equipment transfer to S.P. and B.L.), from Agence Nationale de la Recherche (AAP Prematuration FCS/IDEX Paris Saclay, to A.B. and N.P., France BioImaging infrastructure ANR-10-INBS-04, ANR-11-EQPX-029 to N.P.), from Fondation pour la Recherche sur le Cerveau/Rotary Club de France (B.L.), and from RFBR (project number 20-04-00085 to N.P.M. and E.S.V.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Предметные рубрики: IN-VIVO
PHOTOPROTEIN AEQUORIN
CA2+-REGULATED PHOTOPROTEINS
SPREADING
Аннотация: Considerable efforts have been focused on shifting the wavelength of aequorin Ca2+-dependent blue bioluminescence through fusion with fluorescent proteins. This approach has notably yielded the widely used GFP-aequorin (GA) Ca2+ sensor emitting green light, and tdTomato-aequorin (Redquorin), whose bioluminescence is completely shifted to red, but whose Ca2+ sensitivity is low. In the present study, the screening of aequorin mutants generated at twenty-four amino acid positions in and around EF-hand Ca2+-binding domains resulted in the isolation of six aequorin single or double mutants (AequorinXS) in EF2, EF3, and C-terminal tail, which exhibited markedly higher Ca2+ sensitivity than wild-type aequorin in vitro. The corresponding Redquorin mutants all showed higher Ca2+ sensitivity than wild-type Redquorin, and four of them (RedquorinXS) matched the Ca2+ sensitivity of GA in vitro. RedquorinXS mutants exhibited unaltered thermostability and peak emission wavelengths. Upon stable expression in mammalian cell line, all RedquorinXS mutants reported the activation of the P2Y2 receptor by ATP with higher sensitivity and assay robustness than wt-Redquorin, and one, RedquorinXS-Q159T, outperformed GA. Finally, wide-field bioluminescence imaging in mouse neocortical slices showed that RedquorinXS-Q159T and GA similarly reported neuronal network activities elicited by the removal of extracellular Mg2+. Our results indicate that RedquorinXS-Q159T is a red light-emitting Ca2+ sensor suitable for the monitoring of intracellular signaling in a variety of applications in cells and tissues, and is a promising candidate for the transcranial monitoring of brain activities in living mice.
WOS
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20.

Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Borisova V.V., Frank L.A., Markova S.V., Burakova L.P., Vysotski E.S.
Заглавие : Recombinant Metridia luciferase isoforms: expression, refolding and applicability for in vitro assay
Колич.характеристики :7 с
Коллективы :
Место публикации : Photochem. Photobiol. Sci.: ROYAL SOC CHEMISTRY, 2008. - Vol. 7, Is. 9. - С. 1025-1031. - ISSN 1474-905X, DOI 10.1039/b807271j
Примечания : Cited References: 19. - The work was supported by Bayer AG, by the Russian Foundation for Basic Research grants 05-04-48271 and 06-04-08076, by the joint grant 06-04-89502 of the Russian Foundation for Basic Research and Taiwan National Science Council, and by the "Molecular and Cellular Biology" program of the Russian Academy of Sciences.
Предметные рубрики: BIOLUMINESCENT REPORTER
GAUSSIA LUCIFERASE
CDNA
PROTEINS
CLONING
OVEREXPRESSION
PURIFICATION
MUTAGENESIS
ENZYME
OBELIN
Аннотация: The recombinant coelenterazine-dependent luciferases (isoforms MLuc 164 and MLuc39) from the marine copepod Metridia longa were expressed as inclusion bodies in E. coli cells, dissolved in 6 M guanidinium chloride and folded in conditions developed for proteins containing intramolecular disulfide bonds. One of them (MLuc09) was obtained in an active monomeric form with a high yield. The luciferase bioluminescence is found to be initiated not only by free coelenterazine, but also by Ca2+-dependent coelenterazine-binding protein (CBP) of Renilla muelleri on Ca2+ addition. The use of CBP as a "substrate" provides higher light emission and simultaneously the lower level of background. The high purity MLuc39 can be detected down to attomol with a linear range extending over 5 orders of magnitude. The MLuc39 reveals also a high stability towards heating and chemical modification; the chemically synthesized biotinylated derivatives of the luciferase preserve 35-40% of the initial activity The luciferase applicability as an in vitro bioluminescent reporter is demonstrated in model tandem bioluminescent solid-phase microassay combining the Ca2+-regulated photoprotein obelin and the Metridia luciferase.
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