8 results on '"Artemov AV"'
Search Results
2. [Comparative analysis of methods for purification and concentration of 26S proteasomes isolated from rat liver].
- Author
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Evteeva IN, Starkova TIu, Artemov AV, Zaĭkova IuIa, and Barlev NA
- Subjects
- Animals, Cytoplasm chemistry, Multiprotein Complexes chemistry, Proteasome Endopeptidase Complex chemistry, Proteolysis, Rats, Liver chemistry, Multiprotein Complexes isolation & purification, Proteasome Endopeptidase Complex isolation & purification
- Abstract
The 26S proteasome is a multi-subunit protein complex that consists of the catalytic 20S and regulatory 19S sub-complexes. The most well studied function of proteasomes is specific degradation of proteins. There are several purification schemes for obtaining the preparations of 26S proteasomes. An important step in purification of 26S proteasomes is concentration of the purified material for subsequent analysis of its biochemical functions. In this report we showed that the subunits composition of 26S proteasomes that have been concentrated by the different modes at the latest stage of their preparation is identical. However, the concentrating mode differently affects the functional activity of these complexes.
- Published
- 2013
3. [Comparative structural and functional characteristics of different forms of Saccharomyces cerevisiae red pigment and its synthetic analogue].
- Author
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Amen TP, Mikhaĭlov EV, Alenin VV, Artemov AV, Dement'ev PA, Khodorkovskiĭ MA, Artamonov TO, Kuznetsova IM, Soĭdla TR, and Nevzgliadova OV
- Subjects
- Amino Acids analysis, Binding Sites, Dinitrocresols chemistry, Hydrolysis, Mass Spectrometry, Microscopy, Atomic Force, Molecular Structure, Polymers chemistry, Ribose chemistry, Saccharomyces cerevisiae, Amyloid chemistry, Amyloid drug effects, Insulin chemistry, Insulin Antagonists chemical synthesis, Insulin Antagonists chemistry, Insulin Antagonists pharmacology, Pigments, Biological chemical synthesis, Pigments, Biological chemistry, Pigments, Biological pharmacology
- Abstract
Structural and functional characteristics of the yeast red pigment (product of polymerization of N1-(beta-D-ribofuranosyl)-5-aminoimadazole), isolated from adel 1 mutant cells of Saccharomyces cerevisiae, its deribosylated derivatives (obtained by acid hydrolysis) and its synthetic pigment analogue (product of polymerization of N1-methyl-5-aminoimadazole in vitro) has been obtained. Products of in vitro polymerization were identified using mass spectrometry. The ability of these pigments to inhibit amyloid formation using insulin fibrils was compared. The entire compounds studied were able to interact with amyloids and inhibit their growth. Electron and atomic force microscopy revealed a common feature inherent in the insulin fibrils formed in presence of these compounds--they were merged into conglomerates that were more stable and resistant to the effects of ultrasound in comparison with insulin aggregates grown without pigments. We speculate that all these compounds can cause coalescence of fibrils, partially block their loose ends and, thereby, inhibit the attachment of new monomers to growing fibrils.
- Published
- 2012
4. [The effect of red pigment of Saccharomyces cerevisiae on insulin fibril formation in vitro].
- Author
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Mikhaĭlova EV, Artemov AV, Snigirevskaia ES, Artamonova TO, Khodorkovskiĭ MA, Soĭdla TR, and Nevzgliadova OV
- Subjects
- Amyloid antagonists & inhibitors, Amyloid metabolism, Amyloid ultrastructure, Animals, Benzothiazoles, Biomimetic Materials metabolism, Fluorescence, Fluorescent Dyes analysis, Fluorescent Dyes metabolism, Humans, Hydrogen-Ion Concentration, Hydrolysis, Insulin metabolism, Microscopy, Electron, Microscopy, Electron, Transmission, Prion Diseases metabolism, Prion Diseases pathology, Solutions chemistry, Solutions metabolism, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Thiazoles analysis, Amyloid chemistry, Biomimetic Materials chemistry, Insulin chemistry, Pigments, Biological chemistry, Pigments, Biological metabolism, Pigments, Biological therapeutic use, Prion Diseases drug therapy, Saccharomyces cerevisiae chemistry, Thiazoles metabolism
- Abstract
The effect of the yeast red pigment, the result of polymerization of AIR, and of its low molecular weight derivative (presumably devoid of phosphoribosyl moiety) on the formation of amyloid fibrils in vitro was studied. Both the red pigment and its derivative, the result of acid hydrolysis of the original pigment, were shown to diminish the intensity of amyloid bound Thioflavine T fluorescence. Correlation between the decrease of the intensity of Thioflavine T fluorescence and the concentration of both forms of the red pigment was demonstrated. Both forms were also able to compete with Thioflavine T for amyloid fibrils. Electron microscopy permitted to visualize a drop of fibril size in the case of red pigments presence during their formation.
- Published
- 2011
5. [Comparison of crude lysate pellets of isogenic strains of yeast with different prion composition: identification of a set of prion-associated proteins].
- Author
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Nevzgliadova OV, Artemov AV, Mittenberg AG, Kostyleva EI, Mikhaĭlova EV, Solov'ev KV, Kuznetsova IM, Turoverov KK, and Soĭdla TR
- Subjects
- Blotting, Western, Electrophoresis, Gel, Two-Dimensional methods, Oxidative Stress, Prions isolation & purification, Prions metabolism, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae metabolism, Saccharomyces cerevisiae Proteins isolation & purification, Saccharomyces cerevisiae Proteins metabolism, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization methods, Prions chemistry, Saccharomyces cerevisiae chemistry, Saccharomyces cerevisiae Proteins chemistry
- Abstract
A new approach: comparative analysis of proteins of the pellets of crude cell lysates of isogenic strains of Saccharomyces cerevisiae differing by their prion composition permitted to identify a large group of prion-associated proteins in yeast cells. 2D-electrophoresis followed by MALDI-analysis of a recipient [psi-] strain and of [PSI+] cytoductant led to identification of 35 proteins whose aggregation state responded to a shift of prion(s) content. Approximately half of these proteins belonged to functional groups of chaperones and enzyme involved in glucose metabolism. Notable were also proteins involved in translation, in oxidative stress response and in protein degradation. The data obtained are compared with the results of other groups who used other approaches to detecting proteins involved in prion aggregates.
- Published
- 2010
6. [The effect of red pigment on amyloidization of yeast proteins].
- Author
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Nevzgliadova OV, Artemov AV, Mittenberg AG, Mikhaĭlova EV, Kuznetsova IM, Turoverov KK, and Soĭdla TR
- Subjects
- Amyloid genetics, Down-Regulation, Peptide Synthases genetics, Pigments, Biological genetics, Prions genetics, Protein Binding, Saccharomyces cerevisiae enzymology, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae Proteins genetics, Amyloid metabolism, Peptide Synthases metabolism, Pigments, Biological metabolism, Prions metabolism, Saccharomyces cerevisiae metabolism, Saccharomyces cerevisiae Proteins metabolism
- Abstract
Amyloid bound thioflavine T fluorescence was studied in the lysates of yeast strains carrying mutations in genes ADE1 or ADE2 and accumulating red pigment, a result of polymerization of aminoimidazoleribotide (an intermediate of adenine biosynthesis). The fluorescence is drastically enhanced in the case of cells grown in media containing high concentration of adenine (100 mg/l) that blocks accumulation of red pigment. Blocks at first stages of purine biosynthesis de novo also impede red pigment and lead to the same effect on thioflavine fluorescence. At the same time induction of mutations in genes ADE1 or ADE2 in originally white prototrophic strains leads to considerable drop of fluorescence. A fraction of protein polymers was studied by agarose gel electrophoresis and this permitted to conclude that lowering of fluorescence intensity is indeed connected with the decrease of amyloid amount in cells accumulating red pigment. Model experiments with insulin fibers demonstrate that red pigment binds fibrils and blocks their interaction with Thioflavine T. 2D-electrophoretic comparison of pellet proteins of red and white isogenic strains, followed by MALDI, allowed identification of 23 pigment-dependent proteins. These proteins mostly belong to functional classes of chaperones and proteins, involved in glucose metabolism, closely corresponding to prion-dependent proteins characterized in our previous work. We suppose that, binding amyloid fibrils, red pigment hinders formation of prion aggregates and also, blocking fibril contact with chaperones, impedes prion propagation.
- Published
- 2010
7. [Estimating of changes in the amyloid and prion content of yeast cells].
- Author
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Nevzgliadova OV, Kuznetsova IM, Artemov AV, Mikhaĭlova EV, Turoverov KK, and Soĭdla TR
- Subjects
- Amyloid analysis, Benzothiazoles, Crosses, Genetic, Guanidine, Peptide Termination Factors, Prions metabolism, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae Proteins analysis, Staining and Labeling methods, Thiazoles, Amyloid metabolism, Saccharomyces cerevisiae metabolism, Saccharomyces cerevisiae Proteins metabolism
- Abstract
An attempt was made at estimating the overall amyloid content of yeast cells by treating crude cellular lysates with thioflavin T, the agent specifically staining amyloid fibrils. We demonstrated that overproduction of the yeast chaperone Hsp104p, as well as GuHCI treatment of the [PSI+] cells led both to elimination of the [PSI+] factor and to a stable decrease of the overall amyloid content estimated by intensity of fluorescence (IF) of the thioflavin T. At the same time, overexpression of gene SUP35, coding the protein prionizable to [PSI+], led to generation of [PSI+] clones with higher IF of thioflavin T. Cytoduction in the crosses involving PSI factor leads to considerable enhancement of IF; cytoductants with the nucleus of the recipient [psi-] strain not only got [PSI+] factor from the donor strain but also increased their amyloid content. In these model experiments all treatments modifying one of the yeast prions, [PSI+] factor, led to a predictable shift of IF of thioflavin T that behaved like a cytoplasmic hereditary determinant. The data obtained show that IF of thioflavin T staining gives reliable estimates of cellular amyloid content and that mitotically stable shift of IF after a battery of treatments modifying cellular prion set provides quantitative estimate of the input of prionizable protein molecules to the amyloid pool. The combination of thioflavin staining and prionotropic treatments applied here can be possibly used for future attempts of checking yeast strains for cryptic prions.
- Published
- 2008
8. [Expression of recombinant actin 5C from Drosophila in the methylotrophic yeast Pichia pastoris].
- Author
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Nevzgliadova OV, Artemov AV, Zenin VV, Verkhusha VV, Shavlovskiĭ MM, Povarova OI, Stepanenko OV, Kuznetsova IM, and Turoverov KK
- Subjects
- Actins genetics, Animals, Recombinant Proteins biosynthesis, Actins biosynthesis, Drosophila chemistry, Pichia metabolism, Protein Engineering
- Abstract
A system for actin expression in cells of yeast Pichia pastoris was constructed. Drosophila actin 5C, by 90% homologous to beta-actin of higher eukaryotes, was used as a target protein. To improve the procedures of target protein biosynthesis in yeast cells and of extraction and purification of recombinant actin the fusion protein GFP-actin 5C, having fluorescence protein GFP as a reporter part, was expressed and purified. The dimensions and resistance of yeast cells producing recombinant actin were characterized. It was shown that the size and form of cells depended on the accumulation of recombinant protein. The purified fusion protein was used for obtaining polyclonal antibody for testing recombinant actin.
- Published
- 2007
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