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Formation of the vertical heterogeneity in the Lake Shira ecosystem: The biological mechanisms and mathematical model / A. G. Degermendzhy [et al.] // Aquatic Ecology. - 2002. - Vol. 36, Is. 2. - P271-297, DOI 10.1023/A:1015621508971 . - ISSN 1386-2588
Кл.слова (ненормированные):
Hydrogen sulphide -- Phytoplankton -- Stratification control -- Sulphate-reducing bacteria -- Sulphur cycle -- Vertical model -- biological production -- community structure -- ecosystem modeling -- nutrient cycling -- plankton -- saline lake -- seasonal variation -- vertical distribution -- Russian Federation -- algae -- Arctodiaptomus -- Bacteria (microorganisms) -- Calanoida -- Chlorophyta -- Copepoda -- Crustacea -- Cyanobacteria -- Dictyosphaerium -- Lyngbya -- Lyngbya contorta
Аннотация: Data on the seasonal changes in vertical heterogeneity of the physical-chemical and biological parameters of the thermally stratified Shira Lake ecosystem (Khakasia, Siberia) in 1996-2000 have been analyzed. The interaction mechanisms involving: (1) The plankton populations in aerobic and anaerobic zones, involving the cycling of carbon and sulphur, (2) the primary production limitation (by light and phosphorus) and inhibition (by light), and (3) the kinetic characteristics of plankton populations have been elucidated. A mathematical model of the vertical structure of the lake's plankton populations, based on the ecosystem description and on vertical turbulent diffusion of the matter, has been constructed. The green alga Dictyosphaerium tetrachotomum (Chlorophyta) and the cyanobacterium Lyngbya contorta (Cyanophyta), which dominated the phytoplankton biomass, were taken as oxygen producers. Arctodiaptomus salinus (a calanoid copepod) has been assumed as the main grazer in Shira Lake as it dominated the zooplankton biomass. Four groups of microorganisms involved in the sulphur cycle formation have been distinguished: sulphur, sulphur purple, sulphur green and SRB. H2S is oxidized to sulphate (only the green sulphur bacteria oxidize it to sulphur), and sulphate is reduced to H2S, forming neither sulphur nor its water-soluble compounds. The role of grazing, light and nutrient limitation, in forming the vertical inhomogeneities, particularly in lowering the depth of the maximal cyanobacterial biomass, has been demonstrated. When the model takes into account both light limitation and nutrient limitation of algal growth by P and consumption of algae by crustaceans: (a) in the scenario where the P is formed only by the cycling and decomposition of autochthonous organic matter, both the green algae and cyanobacteria are eliminated; (b) in the scenario involving an additional P flux in the deep water layers the peak of the cyanobacteria is at a depth of 10 m, and its amplitude is close to the one observed in the lake. The position of the peak remains stable owing to the 'double' limitation mechanism: light 'from above' and P 'from below'. Another mechanism responsible for the deep position of the peak of cyanobacteria was analyzed mathematically based on the model involving the experimentally proven assumption of the growth inhibition by light in the epilimnion and the light limitation in the hypolimnion. The main result is: the peak is positioned stable at its depth and does not change with time. The analytical and numerical calculations made for this positioning mechanism yielded the formulae relating the depth of the maximum of algal biomass, the 'width' of the peak base and the peak amplitude and a number of parameters (algae elimination, turbulent diffusion coefficient, sedimentation rate, light extinction coefficient and light intensity). The theoretical curves for the stratification of chemical and biological parameters have been brought in conformity with field observations, e.g. for the different patterns for the peaks, and the biomass maxima of cyanobacteria, purple and green sulphur bacteria, oxygen, and hydrogen sulphide. The calculations revealed that for an adequate assessment of the parameters for the hydrogen sulphide zone it is necessary to introduce flows of allochthonous organic matter. For the first time, theoretically, based on the form of the sulphur distribution curve, the allochthonous input of organic matter and the inflow of hydrogen sulphide from the bottom have been discriminated. The theoretical limit for the depth up to which the hydrogen-sulphide zone can ascend under the impact of allochthonous organic loading, has been determined.

Scopus
Держатели документа:
Institute of Biophysics of SB RAS, Krasnoyarsk 660036, Russian Federation : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Degermendzhy, A.G.; Belolipetsky, V.M.; Zotina, T.A.; Gulati, R.D.

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Growth of dominant zooplankton species feeding on plankton microflora in Lake Shira / T. A. Temerova, A. P. Tolomeyev, A. G. Degermendzhy // Aquatic Ecology. - 2002. - Vol. 36, Is. 2. - P235-243, DOI 10.1023/A:1015607304508 . - ISSN 1386-2588
Кл.слова (ненормированные):
Arctodiaptomus salinus -- Brachionus plicatilis -- Life history -- Reproduction -- Specific growth rate -- diet -- food limitation -- growth rate -- life history -- reproduction -- saline lake -- zooplankton -- Russian Federation -- algae -- Arctodiaptomus -- Bacteria (microorganisms) -- Brachionus -- Brachionus plicatilis -- Calanoida -- Copepoda -- Cyanobacteria -- Invertebrata -- Rotifera
Аннотация: Batch cultures and continuous flow cultures were used to study the growth rates of zooplankton species from Shira lake, the rotifer Brachionus plicatilis Muller and calanoid copepod Arctodiaptomus salinus Daday, which were fed on phytoplankton and bacterioplankton from the lake. Analyses of the birth and survival rates were used to demonstrate that the lake phytoplankton, consisting mostly of cyanobacteria and diatomaceous algae, is inadequotes for optimal realisation of the reproductive potential of B. plicatilis when compared with the bacterial diet. The study revealed that the kinetic growth characteristics of the two zooplankters were similar: B. plicatilis rmax, 0.120 d-1; S0, 0.253; and Ks, 0.114 mg dry mass l-1; and for A. salinus rmax, 0.129 d-1; S0, 0.240; and Ks, 0.171 mg dry mass l-1. Fluctuations in natural food concentration reduced the growth rate of both species. Even though the threshold concentration of food for B. plicatilis and A. salinus were quite similar, the copepods were less sensitive to food limitation.

Scopus
Держатели документа:
Institute of Biophysics of SB RAS, Krasnoyarsk 660036, Russian Federation : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Temerova, T.A.; Tolomeyev, A.P.; Degermendzhy, A.G.

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Effect of Water Temperature on the Content of Essential Polyunsaturated Fatty Acids in Freshwater Zooplankton / M. I. Gladyshev [et al.] // Doklady Biochemistry and Biophysics. - 2011. - Vol. 437, Is. 1. - P57-59, DOI 10.1134/S1607672911020013 . - ISSN 1607-6729
Кл.слова (ненормированные):
docosahexaenoic acid -- fresh water -- icosapentaenoic acid -- unsaturated fatty acid -- water -- animal -- article -- chemistry -- copepod -- environmental monitoring -- metabolism -- temperature -- zooplankton -- Animals -- Copepoda -- Docosahexaenoic Acids -- Eicosapentaenoic Acid -- Environmental Monitoring -- Fatty Acids, Unsaturated -- Fresh Water -- Temperature -- Water -- Zooplankton

Scopus
Держатели документа:
Institute of Biophysics, Siberian Branch of Russian Academy of Sciences, Akademgorodok, Krasnoyarsk 660036, Russian Federation
Scientific-Production Center of the National Academy of Sciences of Belarus for Bioresources, Minsk, Belarus
Institute of Biology of Komi Scientific Center, Ural Branch, Russian Academy of Sciences, Syktyvkar 167000, Russian Federation
Siberian Federal University, Krasnoyarsk 660041, Russian Federation
Kamchatka Research Institute of Fisheries and Oceanography, Naberezhnaya 18, Petropavlovsk-Kamchatskii 683602, Russian Federation : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Gladyshev, M.I.; Semenchenko, V.P.; Dubovskaya, O.P.; Fefilova, E.B.; Makhutova, O.N.; Buseva, Z.F.; Sushchik, N.N.; Baturina, M.A.; Razlutskij, V.I.; Lepskaya, E.V.; Kalacheva, G.S.

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Chemical interactions between two dominant species in Lake Shira: Gammarus lacustris sars (Crustacea: Amphipoda) and Arctodiaptomus salinus Daday (Crustacea: Copepoda) / M. V. Gubanov, E. S. Zadereev, A. G. Degermenzhy // Doklady Biological Sciences. - 2010. - Vol. 434, Is. 1. - P318-321, DOI 10.1134/S0012496610050078 . - ISSN 0012-4966
Кл.слова (ненормированные):
fresh water -- pheromone -- Amphipoda -- animal -- article -- copepod -- drug effect -- ecosystem -- escape behavior -- physiology -- Russian Federation -- Amphipoda -- Animals -- Copepoda -- Ecosystem -- Escape Reaction -- Fresh Water -- Pheromones -- Russia -- Amphipoda -- Arctodiaptomus salinus -- Copepoda -- Crustacea -- Gammarus -- Gammarus lacustris

Scopus
Держатели документа:
Siberian Federal University, pr. Svobodnyi 79, Krasnoyarsk 660041, Russian Federation
Institute of Biophysics, Siberian Branch, Russian Academy of Sciences, Academgorodok, Krasnoyarsk 660036, Russian Federation : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Gubanov, M.V.; Zadereev, E.S.; Degermenzhy, A.G.

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The vertical distribution of zooplankton in brackish meromictic lake with deep-water chlorophyll maximum / Y. S. Zadereev, A. P. Tolomeyev // Hydrobiologia. - 2007. - Vol. 576, Is. 1. - P69-82, DOI 10.1007/s10750-006-0294-x . - ISSN 0018-8158
Кл.слова (ненормированные):
Anoxic hypolimnion -- Deep water chlorophyll maximum -- Meromictic lake -- Stratification -- Vertical distribution -- Zooplankton -- Algae -- Chlorophyll -- Growth kinetics -- Lakes -- Saline water -- Anoxic hypolimnion -- Deep water chlorophyll maximum -- Meromictic lake -- Stratification -- Vertical distribution -- Zooplanktons -- Biodiversity -- biomass -- brackish water -- chlorophyll -- green alga -- meromictic lake -- reproduction -- stratification -- vertical distribution -- zooplankton -- Eurasia -- Khakassia -- Lake Shira -- Russian Federation -- Arctodiaptomus salinus -- Brachionus plicatilis -- Chlorophyta -- Copepoda -- Hexarthra oxiuris -- Rotifera
Аннотация: We examined the dynamics of the vertical stratification of physical, chemical and biological factors in a brackish meromictic lake with a deep-water chlorophyll maximum (Shira Lake, Russia, Khakasia) during the growing season and estimated how the vertical distribution of these factors influences the vertical distribution of the zooplankton community. The vertical distribution of zooplankton was restricted by the anoxic hypolimnion. Nauplii and younger copepodides (C1-C3) of the copepod, Arctodiaptomus salinus, and the rotifer, Brachionus plicatilis, were found in the upper warm waters. During summer stratification the maximum of A. salinus biomass, which consisted mainly of older copepodides (C4-C5) and females, was associated with the deep-water maximum of biomass of green algae, which are the preferred diet for this species. The vertical distribution of the rotifer Hexarthra oxiuris was bimodal with numerical peaks in the epi- and hypolimnion. Reproduction peaks of dominant species were separated in time. The reproduction peak of A. salinus was at the beginning of summer when A. salinus constituted up to 99% of total zooplankton biomass. The development of rotifers was detected after the reproduction peak of A. salinus when the biomass of rotifers reached 50% of total zooplankton biomass. В© 2007 Springer Science+Business Media B.V.

Scopus
Держатели документа:
Institute of Biophysics SB RAS, Akademgorodok, Krasnoyarsk 660036, Russian Federation : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Zadereev, Y.S.; Tolomeyev, A.P.

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High-active truncated luciferases of copepod Metridia longa and their characterization as secreted reporters inmammalian cells / S. V. Markova, L. P. Burakova, E. S. Vysotski // Luminescence. - 2012. - Vol. 27, Is. 2. - P138-139. - Cited References: 1 . - ISSN 1522-7235

РУБ Biochemistry & Molecular Biology

Держатели документа:
[Markova, Svetlana V.
Burakova, Ludmila P.
Vysotski, Eugene S.] Inst Biophys SB RAS, Photobiol Lab, Krasnoyarsk 660036, Russia
[Markova, Svetlana V.
Burakova, Ludmila P.
Vysotski, Eugene S.] Siberian Fed Univ, Krasnoyarsk 660041, Russia
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50
Доп.точки доступа:
Markova, S.V.; Burakova, L.P.; Vysotski, E.S.

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High-active truncated luciferase of copepod Metridia longa / S. V. Markova, L. P. Burakova, E. S. Vysotski // Biochem. Biophys. Res. Commun. - 2012. - Vol. 417, Is. 1. - P98-103, DOI 10.1016/j.bbrc.2011.11.063. - Cited References: 31. - This study was supported by the Grants 16.512.11.2141 and 64987.2010.4 of the Ministry of Education and Science of Russian Federation. . - ISSN 0006-291X

РУБ Biochemistry & Molecular Biology + Biophysics
Рубрики:
COELENTERAZINE-BINDING PROTEIN
   REPORTER-GENE-EXPRESSION
   RENILLA LUCIFERASE
   GAUSSIA LUCIFERASE
   LIGHT-EMITTER
   IN-VIVO
   BIOLUMINESCENCE
   PHOTOPROTEINS
   CDNA
   SUBSTRATE
Кл.слова (ненормированные):
Bioluminescence -- Coelenterazine -- Mammalian expression -- Secretion
Аннотация: The technology of real-time imaging in living cells is crucial for understanding of intracellular events. For this purpose, bioluminescent reporters have been introduced as sensitive and convenient tools. Metridia luciferase (MLuc) from the copepod Metridia longa is a coelenterazine-dependent luciferase containing a natural signal peptide for secretion. We report the high-active MLuc mutants with deletion of the N-terminal variable part of amino acid sequence. The MLuc variants were produced in Escherichia coil cells, converted to an active protein, and characterized. We demonstrate that the truncated MLucs have significantly increased bioluminescent activity as against the wild type enzyme but substantially retain other properties. One of the truncated variants of MLuc was transiently expressed in HEK 293 cells. The results clearly suggest that the truncated Metridia luciferase is well suited as a secreted reporter ensuring higher detection sensitivity in comparison with a wild type enzyme. (C) 2011 Elsevier Inc. All rights reserved.

Держатели документа:
[Vysotski, Eugene S.] Russian Acad Sci, Photobiol Lab, Inst Biophys, Siberian Branch, Krasnoyarsk 660036, Russia
Siberian Fed Univ, Dept Biophys, Krasnoyarsk 660041, Russia
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Markova, S.V.; Burakova, L.P.; Vysotski, E.S.

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Expression, purification and characterization of the secreted luciferase of the copepod Metridia longa from Sf9 insect cells [Text] / G. A. Stepanyuk [et al.] // Protein Expr. Purif. - 2008. - Vol. 61, Is. 2. - P142-148, DOI 10.1016/j.pep.2008.05.013. - Cited References: 34. - This work was supported by the National Institutes of Health (Grant 1P50 GM62407), University of Georgia Research Foundation and Georgia Research Alliance, the Russian Foundation for Basic Research and Taiwan National Science Council (Grant 06-0489502) and the program for "Molecular and Cellular Biology" of Russian Academy of Sciences. . - ISSN 1046-5928

РУБ Biochemical Research Methods + Biochemistry & Molecular Biology + Biotechnology & Applied Microbiology
Рубрики:
VARGULA-HILGENDORFII LUCIFERASE
   CRYSTAL-STRUCTURE
   RENILLA-RENIFORMIS
   GAUSSIA LUCIFERASE
   BIOLUMINESCENT REPORTER
   OBELIN BIOLUMINESCENCE
   ANGSTROM RESOLUTION
   MAMMALIAN-CELLS
   GENE-EXPRESSION
   IN-VIVO
Аннотация: Metridia luciferase is a secreted luciferase from a marine copepod and uses coelenterazine as a substrate to produce a blue bioluminescence This luciferase has been successfully applied as a bioluminescent reporter in mammalian cells. The main advantage of secreted luciferase as a reporter is the capability of measuring intracellular events without destroying the cells or tissues and this property is well suited for development of high throughput screening technologies. However because Metridia luciferase is a Cys-rich protein, Escherichia coli expression systems produce an incorrectly folded protein, hindering its biochemical characterization and application for development of in vitro bioluminescent assays. Here we report the successful expression of Metridia luciferase with its signal peptide for secretion, in insect (Sf9) cells using the baculovirus expression system. Functionally active luciferase secreted by insect cells into the culture media has been efficiently purified with a yield of high purity protein of 2-3mg/L This Metridia luciferase expressed in the insect cell system is a monomeric protein showing 3.5-fold greater bioluminescence activity than luciferase expressed and purified from E. coli. The near coincidence of the experimental mass of Metridia luciferase purified from insect cells with that calculated from amino acid sequence. indicates that luciferase does not undergo post-translational modifications such as phosphorylation or glycosylation and also, the cleavage site of the signal peptide for secretion is at VQA-KS, as predicted from sequence analysis. (c) 2008 Elsevier Inc. All rights reserved.

Держатели документа:
[Stepanyuk, Galina A.
Markova, Svetlana V.
Vysotski, Eugene S.] Russian Acad Sci, Photobiol Lab, Inst Biophys, Siberian Branch, Krasnoyarsk 660036, Russia
[Stepanyuk, Galina A.
Xu, Hao
Wu, Chia-Kuei
Lee, John
Vysotski, Eugene S.
Wang, Bi-Cheng] Univ Georgia, Dept Biochem & Mol Biol, Athens, GA 30602 USA
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Stepanyuk, G.A.; Xu, H...; Wu, C.K.; Markova, S.V.; Lee, J...; Vysotski, E.S.; Wang, B.C.

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Recombinant Metridia luciferase isoforms: expression, refolding and applicability for in vitro assay [Text] / V. V. Borisova [et al.] // Photochem. Photobiol. Sci. - 2008. - Vol. 7, Is. 9. - P1025-1031, DOI 10.1039/b807271j. - 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. . - ISSN 1474-905X

РУБ Biochemistry & Molecular Biology + Biophysics + Chemistry, Physical
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.

Держатели документа:
[Borisova, Vasillisa V.
Frank, Ludmila A.
Markova, Svetlana V.
Burakova, Ludmilla P.
Vysotski, Eugene S.] Russian Acad Sci, Inst Biophys, Photobiol Lab, Siberian Branch, Krasnoyarsk 660036, Russia
[Frank, Ludmila A.] Siberian Fed Univ, Krasnoyarsk 660041, Russia
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Borisova, V.V.; Frank, L.A.; Markova, S.V.; Burakova, L.P.; Vysotski, E.S.

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10.

Cloning and expression of cDNA for a luciferase from the marine copepod Metridia longa - A novel secreted bioluminescent reporter enzyme [Text] / S. V. Markova [et al.] // J. Biol. Chem. - 2004. - Vol. 279, Is. 5. - P3212-3217, DOI 10.1074/jbc.M309639200. - Cited References: 37 . - ISSN 0021-9258

РУБ Biochemistry & Molecular Biology
Рубрики:
VARGULA-HILGENDORFII LUCIFERASE
   GREEN FLUORESCENT PROTEIN
   GENE-EXPRESSION
   FIREFLY LUCIFERASE
   PROMOTER ACTIVITY
   MAMMALIAN-CELLS
   RECEPTOR
   CANCER
   PHOTOPROTEINS
   LUMINESCENCE
Аннотация: Metridia longa is a marine copepod from which a blue bioluminescence originates as a secretion from epidermal glands in response to various stimuli. We demonstrate that Metridia luciferase is specific for coelenterazine to produce blue light (lambda(max)=480 nm). Using an expression cDNA library and functional screening, we cloned and sequenced the cDNA encoding the Metridia luciferase. The cDNA is an 897-bp fragment with a 656-bp open reading frame, which encodes a 219-amino acid polypeptide with a molecular weight of 23,885. The polypeptide contains an N-terminal signal peptide of 17 amino acid residues for secretion. On expression of the Metridia luciferase gene in mammalian Chinese hamster ovary cells the luciferase is detected in the culture medium confirming the existence of a naturally occurring signal peptide for secretion in the cloned luciferase. The novel secreted luciferase was tested in a practical assay application in which the activity of A2a and NPY2 G-protein-coupled receptors was detected. These results clearly suggest that the secreted Metridia luciferase is well suited as a reporter for monitoring gene expression and, in particular, for the development of novel ultra-high throughput screening technologies.

Держатели документа:
Russian Acad Sci, Siberian Branch, Inst Biophys, Photobiol Lab, Krasnoyarsk 660036, Russia
Bayer AG, Pharma Res Mol Screening Technol, D-42096 Wuppertal, Germany
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Markova, S.V.; Golz, S...; Frank, L.A.; Kalthof, B...; Vysotski, E.S.

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11.

The production of highly active recombinant 16.5 kDa-isoform of Metridia longa luciferase, the smallest copepod luciferase [Text] / M. . Larionova [et al.] // Luminescence. - 2014. - Vol. 29. - P79-80. - Cited References: 3 . - ISSN 1522-7235. - ISSN 1522-7243
Рубрики:
EXPRESSION

WOS
Держатели документа:
[Larionova, Marina
Markova, Svetlana
Burakova, Ludmila
Vysotski, Eugene] Russian Acad Sci, Inst Biophys, Photobiol Lab, Siberian Branch, Krasnoyarsk, Russia
[Larionova, Marina] Siberian Fed Univ, Inst Fundamental Biol & Biotecnol, Lab Bioluminescence Biotechnol, Krasnoyarsk, Russia
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Larionova, M...; Markova, S...; Burakova, L...; Vysotski, E...

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12.

The smallest natural high-active luciferase: Cloning and characterization of novel 16.5-kDa luciferase from copepod Metridia longa [Text] / S. V. Markova [et al.] // Biochem. Biophys. Res. Commun. - 2015. - Vol. 457, Is. 1. - P77-82, DOI 10.1016/j.bbrc.2014.12.082. - Cited References:20. - The cloning of cDNA encoding MLuc7 luciferase of M. longa was supported by Bayer AG (Germany); all other studies - by the grant 14-14-01119 of the Russian Science Foundation. We declare that authors have no conflict of interest. . - ISSN 0006-291X. - ISSN 1090-2104

РУБ Biochemistry & Molecular Biology + Biophysics
Рубрики:
CDNA CLONING
   SECRETED LUCIFERASE
   ESCHERICHIA-COLI
   EXPRESSION
Кл.слова (ненормированные):
Bioluminescence -- Coelenterazine -- Copepod luciferase -- Mammalian -- expression -- Real-time imaging
Аннотация: Coelenterazine-dependent copepod luciferases containing natural signal peptide for secretion are a very convenient analytical tool as they enable monitoring of intracellular events with high sensitivity, without destroying cells or tissues. This property is well suited for application in biomedical research and development of cell-based assays for high throughput screening. We report the cloning of cDNA gene encoding a novel secreted non-allelic 16.5-kDa isoform (MLuc7) of Metridia longa luciferase, which, in fact, is the smallest natural luciferase of known for today. Despite the small size, isoform contains 10 conservative Cys residues suggesting the presence of up to 5 S-S bonds. This hampers the efficient production of functionally active recombinant luciferase in bacterial expression systems. With the use of the baculovirus expression system, we produced substantial amounts of the proper folded MLuc7 luciferase with a yield of similar to 3 mg/L of a high purity protein. We demonstrate that MLuc7 produced in insect cells is highly active and extremely thermostable, and is well suited as a secreted reporter when expressed in mammalian cells ensuring higher sensitivity of detection as compared to another Metridia luciferase isoform (MLuc164) which is widely employed in real-time imaging. (C) 2014 Elsevier Inc. All rights reserved.

WOS
Держатели документа:
Russian Acad Sci, Inst Biophys, Photobiol Lab, Siberian Branch, Krasnoyarsk, Russia.
Siberian Fed Univ, Chair Biophys, Krasnoyarsk, Russia.
ИБФ СО РАН

Доп.точки доступа:
Markova, Svetlana V.; Larionova, Marina D.; Burakova, Ludmila P.; Vysotski, Eugene S.; Bayer AG (Germany); Russian Science Foundation [14-14-01119]

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13.

The novel extremely psychrophilic luciferase from Metridia longa: Properties of a high-purity protein produced in insect cells / M. D. Larionova, S. V. Markova, E. S. Vysotski // Biochem. Biophys. Res. Commun. - 2017. - Vol. 483, Is. 1. - P772-778, DOI 10.1016/j.bbrc.2016.12.067. - 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. . - ISSN 0006-291X. - ISSN 1090-2104

РУБ Biochemistry & Molecular Biology + Biophysics
Рубрики:
CDNA CLONING
EXPRESSION
ENZYME
Кл.слова (ненормированные):
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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Fed Res Ctr Krasnoyarsk Sci Ctr SB RAS, Inst Biophys SB RAS, Photobiol Lab, Krasnoyarsk, Russia.
Siberian Fed Univ, Krasnoyarsk, Russia.

Доп.точки доступа:
Larionova, Marina D.; Markova, Svetlana V.; Vysotski, Eugene S.; Bayer AG (Germany); Russian Academy of Sciences [01201351504]; RFBR; Government of Krasnoyarsk Territory [16-44-242099]

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14.

Tyr72 and Tyr80 are Involved in the Formation of an Active Site of a Luciferase of Copepod Metridia longa / M. D. Larionova, S. V. Markova, E. S. Vysotski // Photochem. Photobiol. - 2017. - Vol. 93, Is. 2. - P503-510, DOI 10.1111/php.12694. - Cited References:41. - This work was supported by the grant 14-14-01119 of the Russian Science Foundation. . - ISSN 0031-8655. - ISSN 1751-1097

РУБ Biochemistry & Molecular Biology + Biophysics
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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Krasnoyarsk Sci Ctr SB RAS, Inst Biophys SB RAS, Fed Res Ctr, Photobiol Lab, Krasnoyarsk, Russia.
Siberian Fed Univ, Chair Biophys, Krasnoyarsk, Russia.

Доп.точки доступа:
Larionova, Marina D.; Markova, Svetlana V.; Vysotski, Eugene S.; Russian Science Foundation [14-14-01119]

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15.

The disulfide-rich Metridia luciferase refolded from E. coli inclusion bodies reveals the properties of a native folded enzyme produced in insect cells / S. V. Markova [et al.] // J. Photochem. Photobiol. B-Biol. - 2017. - Vol. 175. - P51-57, DOI 10.1016/j.jphotobiol.2017.08.024. - 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). . - ISSN 1011-1344

РУБ Biochemistry & Molecular Biology + Biophysics
Рубрики:
GAUSSIA-PRINCEPS LUCIFERASE
   ESCHERICHIA-COLI
   EXPRESSION
   PROTEIN
Кл.слова (ненормированные):
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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RAS, Krasnoyarsk Sci Ctr SB, Fed Res Ctr, Photobiol Lab,Inst Biophys SB, Krasnoyarsk, Russia.
Siberian Fed Univ, Krasnoyarsk, Russia.

Доп.точки доступа:
Markova, Svetlana V.; Larionova, Marina D.; Gorbunova, Darya A.; Vysotski, Eugene S.; RFBR; Government of Krasnoyarsk Territory [16-44-242099]; Russian Academy of Sciences [0356-2016-0712]

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16.

Effects of water column processes on the use of sediment traps to measure zooplankton non-predatory mortality: a mathematical and empirical assessment / O. P. Dubovskaya [et al.] // J. Plankton Res. - 2018. - Vol. 40, Is. 1. - P91-106, DOI 10.1093/plankt/fbx066. - Cited References:49. - This work was a part of the joint German-Russian Project "Mortality of Zooplankton in lake ecosystems and its potential contribution to carbon mineralization in pelagic zone" supported by the German Research Foundation (DFG no. GR-1540/29-1) and the Russian Foundation for Basic Research (RFBR no. 16-54-12048). The work also was partly supported by Russian Federal Tasks of Fundamental Research (project no. 51.1.1) and by grant (no. 9249.2016.5) from the RF President Council on Grants for leading RF scientific schools. . - ISSN 0142-7873. - ISSN 1464-3774

РУБ Marine & Freshwater Biology + Oceanography
Рубрики:
NONCONSUMPTIVE MORTALITY
CRUSTACEAN ZOOPLANKTON
VERTICAL-DISTRIBUTION
Кл.слова (ненормированные):
zooplankton -- Arctodiaptomus salinus -- non-predatory mortality -- sediment -- trap -- carcasses -- stratified lake
Аннотация: Zooplankton populations can at times suffer mass mortality due to non-predatory mortality (NPM) factors, and the resulting carcasses can be captured by sediment traps to estimate NPM rate. This approach assumes sinking to be the primary process in removing carcasses, but in reality, carcasses can also be removed by ingestion, turbulent mixing and microbial degradation in the water column. We present mathematical formulations to calculate NPM from sediment trap data by accounting for carcass removal by processes in addition to sinking, and demonstrate their application in a study in Lake Shira, Russia. Carcass abundance of the major calanoid copepod Arctodiaptomus salinus decreased with depth, indicating the effect of carcass removal from the water column. The estimated NPM values (0.0003-0.103 d(-1)) were comparable with previously reported physiological death rates. We further used independent data to partition carcass removal due to detritivory, turbulent mixing and microbial degradation. Estimated ingestion by the amphipod Gammarus lacustris could account for the disappearance of copepod carcasses above the traps. Wind-driven turbulence could also extend the carcass exposure time to microbial degradation. Collectively, these water column processes would facilitate the remineralization of carcasses in the water column, and diminish the carcass carbon flux to the benthos.

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Держатели документа:
Russian Acad Sci, Siberian Branch, Krasnoyarsk Sci Ctr, Inst Biophys,Fed Res Ctr,Dept Expt Hydroecol, 50-50 Akademgorodok, Krasnoyarsk 660036, Russia.
Siberian Fed Univ, Inst Fundamental Biol & Biotechnol, 79 Svobodny Ave, Krasnoyarsk 660041, Russia.
Leibniz Inst Freshwater Ecol & Inland Fisherie, Dept Ecohydrol, Muggelseedamm 310, D-12587 Berlin, Germany.
Natl Acad Sci Belarus Bioresources, Pract Ctr, Dept Hydrobiol, 27 Acad Skaya St, Minsk 220072, Byelarus.
Swansea Univ, Dept Biosci, Singleton Pk, Swansea SA2 8PP, W Glam, Wales.

Доп.точки доступа:
Dubovskaya, Olga P.; Tolomeev, Aleksandr P.; Kirillin, Georgiy; Buseva, Zhanna; Tang, Kam W.; Gladyshev, Michail I.; German Research Foundation (DFG) [GR-1540/29-1]; Russian Foundation for Basic Research (RFBR) [16-54-12048]; Russian Federal Tasks of Fundamental Research [51.1.1]; RF President Council on Grants for leading RF scientific schools [9249.2016.5]

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17.

Bioluminescent and structural features of native folded Gaussia luciferase / M. D. Larionova, S. V. Markova, E. S. Vysotski // J. Photochem. Photobiol. B Biol. - 2018. - Vol. 183. - P309-317, DOI 10.1016/j.jphotobiol.2018.04.050 . - ISSN 1011-1344
Кл.слова (ненормированные):
Bioluminescence -- Coelenterazine -- Copepod luciferase -- Halophilic enzyme -- Kinetic cooperativity
Аннотация: The secreted luciferases responsible for light emission of marine copepods have gained popularity for being used in noninvasive imaging of intracellular events. The secreted luciferase of copepod Gaussia princeps is a one-subunit protein catalyzing coelenterazine oxidation to emit blue light. It consists of the N-terminal variable part that bears a signal peptide for secretion and the C-terminal catalytic domain containing ten highly conserved Cys residues supposing the existence of up to five S–S bonds. Despite wide application of Gaussia luciferase in biomedical research, its biochemical properties are still insufficiently studied due to the general problem of obtaining the proper folded Cys-rich proteins in bacterial cells. Here we report the properties of the proper folded Gaussia luciferase produced in insect cells using baculovirus expression system. This high purity luciferase reveals the highest activity at 15–20 °C but retains only ~20% activity at 37 °C that may hamper its application for in vivo assays. The maximum of bioluminescent activity of GpLuc is found at NaCl concentrations in the range of 1.0–1.5 M and, furthermore, a high NaCl concentration enhances luciferase stability to thermal denaturation, i.e. Gaussia luciferase displays the features characteristic of halophilic enzymes. The studies on bioluminescence kinetics at different coelenterazine concentrations obviously show a positive cooperativity of Gaussia luciferase with coelenterazine (Hill coefficient – 1.8 ± 0.2; K0.5–2.14 ± 0.17 ?M). We suggest this effect to be rather due to the so-called kinetic cooperativity conditioned by conformational changes in response to substrate binding than to the presence of two catalytic sites. © 2018 Elsevier B.V.

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Держатели документа:
Photobiology Laboratory, Institute of Biophysics SB RAS, Federal Research Center “Krasnoyarsk Science Center SB RAS”, Krasnoyarsk, Russian Federation
Siberian Federal University, Krasnoyarsk, Russian Federation

Доп.точки доступа:
Larionova, M. D.; Markova, S. V.; Vysotski, E. S.

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18.

Luminescent properties and application of luciferase isoforms from copepod Metridia longa / M. D. Larionova, S. V. Markova, E. S. Vysotski // FEBS Open Bio. - 2018. - Vol. 8. - P166-166. - Cited References:0. - These studies were funded by RFBR and Government of Krasnoyarsk Territory according to the research project No. 16-44-242099. . - ISSN 2211-5463

РУБ Biochemistry & Molecular Biology

WOS
Держатели документа:
Russian Acad Sci, Inst Biophys, Siberian Branch, Krasnoyarsk, Russia.
Siberian Fed Univ, Krasnoyarsk, Russia.
Доп.точки доступа:
Larionova, M. D.; Markova, S. V.; Vysotski, E. S.; RFBR; Government of Krasnoyarsk Territory [16-44-242099]

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19.

Nonspecific stress response to temperature increase in Gammarus lacustris Sars with respect to oxygen-limited thermal tolerance concept / K. Vereshchagina [et al.] // PeerJ. - 2018. - Vol. 6. - Ст. e5571, DOI 10.7717/peerj.5571. - Cited References:49. - The study was carried out with the main financial support of Russian Science Foundation grant 17-14-01063, with the partial financial support of Russian Foundation for Basic Research grants 16-34-00687, 16-34-60060, 17-34-50012, the base part of Goszadanie project 6.9654.2017/8.9, joint program of DAAD and Ministry of education and Science M. Lomonosov (6.12735.2018/12.2) and Lake Baikal Foundation (FOB_02-3/05). There was no additional external funding received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. . - ISSN 2167-8359

РУБ Multidisciplinary Sciences
Рубрики:
COPEPOD TIGRIOPUS-JAPONICUS
SHUNET SOUTH SIBERIA
HEAT-SHOCK PROTEINS
Кл.слова (ненормированные):
Gammarus lacustris -- Heat shock proteins 70 (HSP70) -- Nonspecific cellular -- stress-response (NCSR) -- Lactate dehydrogenase -- Diene conjugates -- Schiff -- bases -- Triene conjugates
Аннотация: The previously undescribed dynamics of the heat shock protein HSP70 and subsequent lipid peroxidation products have been assessed alongside lactate dehydrogenase activity for Gammarus lacustris Sars, an amphipod species from the saltwater Lake Shira (Republic of Khakassia). Individuals were exposed to a gradual temperature increase of 1 degrees C/hour (total exposure duration of 26 hours) starting from the mean annual temperature of their habitat (7 degrees C) up to 33 degrees C. A complex of biochemical reactions occurred when saltwater G. lactustris was exposed to the gradual changes in temperature. This was characterized by a decrease in lactate dehydrogenase activity and the launching of lipid peroxidation. The HSP70 level did not change significantly during the entire experiment. In agreement with the concept of oxygen-limited thermal tolerance, an accumulation of the most toxic lipid peroxides (triene conjugates and Schiff bases) in phospholipids occurred at the same time and temperature as the accumulation of lactate. The main criterion overriding the temperature threshold was, therefore, the transition to anaerobiosis, confirmed by the elevated lactate levels as observed in our previous associated study, and by the development of cellular stress, which was expressed by an accumulation of lipid peroxidation products. An earlier hypothesis, based on freshwater individuals of the same species, has been confirmed whereby the increased thermotolerance of G. lacustris from the saltwater lake was caused by differences in energy metabolism and energy supply of nonspecific cellular stress-response mechanisms. With the development of global climate change, these reactions could be advantageous for saltwater G. lacustris. The studied biochemical reactions can be used as biomarkers for the stress status of aquatic organisms when their habitat temperature changes.

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Держатели документа:
Irkutsk State Univ, Inst Biol, Irkutsk, Russia.
Baikal Res Ctr, Irkutsk, Russia.
Belarusian State Univ, Int Sakharov Environm Inst, Minsk, BELARUS.
SB RAS, Inst Biophys, Krasnoyarsk Res Ctr, Krasnoyarsk, Russia.
Siberian Fed Univ, Krasnoyarsk, Russia.

Доп.точки доступа:
Vereshchagina, Kseniya; Kondrateva, Elizaveta; Axenov-Gribanov, Denis; Shatilina, Zhanna; Khomich, Andrey; Bedulina, Daria; Zadereev, Egor; Timofeyev, Maxim; Russian Science Foundation [17-14-01063]; Russian Foundation for Basic Research [16-34-00687, 16-34-60060, 17-34-50012]; Goszadanie project joint program of DAAD [6.9654.2017/8.9]; Ministry of education and Science M. Lomonosov [6.12735.2018/12.2]; Lake Baikal Foundation [FOB_02-3/05]

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20.

Shining Light on the Secreted Luciferases of Marine Copepods: Current Knowledge and Applications. / S. V. Markova, M. D. Larionova, E. S. Vysotski // Photochemistry and photobiology. - 2018, DOI 10.1111/php.13077 . - ISSN 1751-1097
Аннотация: Copepod luciferases - a family of small secretory proteins of 18.4-24.3 kDa, including a signal peptide, are responsible for bright secreted bioluminescence of some marine copepods. The copepod luciferases use coelenterazine as a substrate to produce blue light in a simple oxidation reaction without any additional cofactors. They do not share sequence or structural similarity with other identified bioluminescent proteins including coelenterazine-dependent Renilla and Oplophorus luciferases. The small size, strong luminescence activity and high stability, including thermostability, make secreted copepod luciferases very attractive candidates as reporter proteins which are particularly useful for nondisruptive reporter assays and for high-throughput format. The most known and extensively investigated representatives of this family are the first cloned GpLuc and MLuc luciferases from copepods Gaussia princeps and Metridia longa, respectively. Immediately after cloning these homologous luciferases were successfully applied as bioluminescent reporters in vivo and in vitro, and since then the scope of their applications continues to grow. This review is an attempt to systemize and critically evaluate the data scattered through numerous articles regarding the main structural features of copepod luciferases, their luminescent and physicochemical properties. We also review the main trends of their application as bioluminescent reporters in cell and molecular biology. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

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Держатели документа:
Photobiology Laboratory, Institute of Biophysics SB RAS, Federal Research Center "Krasnoyarsk Science Center SB RAS", Krasnoyarsk, 660036, Russia.
Siberian Federal University, Krasnoyarsk, 660041, Russia.
N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia, Moscow, 115478, Russia.

Доп.точки доступа:
Markova, Svetlana V.; Larionova, Marina D.; Vysotski, Eugene S.

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