Anatoliy I. Miroshnikov

Education

PeriodCountry, cityEducation institutionAdditional info
1957–1963 Russia, Moscow Moscow Institute of Fine Chemical Technology, dep. of the Fine Chemical Technology MS in chemistry
1968 Russia, Moscow Institute of Chemistry of the Natural Compounds of the USSR AS PhD in chemistry
1981 Russia, Moscow Moscow Institute of Fine Chemical Technology, dep. of the Fine Chemical Technology Assosiated professor
1989 Russia, Moscow NPO "VILR" Ministry of medical and microbiological industry of the USSR DSc in chemistry

Selected publications

  1. Romanov V.P., Kostromina T.I., Miroshnikov A.I., Feofanov S.A. (2017). Preparative method for obtaining recombinant human interferon α2b from inclusion bodies of Escherichia coli. Russ. J. Bioorgan. Chem. 42 (6), 631–637 [+]

    A simple, easily reproducible, and scalable method for obtaining recombinant human interferon α2b from Escherichia coli inclusion bodies has been elaborated. It involves the following steps: preparation of producer cell biomass, isolation and washing of inclusion bodies, their dissolution with protein refolding, SP Sepharose chromatography, and DEAE Sepharose chromatography. According to the results of gel electrophoresis and reversed-phase HPLC, the purity of the protein obtained exceeds 95%.

    ID:2100
  2. Skoblov M.Y., Shibanova E.D., Kovaleva E.V., Bairamashvili D.I., Skoblov Y.S., Miroshnikov A.I. (2010). DNA Assay for Recombinant Pharmaceutical Substances Using the Real_Time PCR Technique. Russ. J. Bioorgan. Chem. 36 (1), 104–108 ID:266
  3. Безуглов В.В., Грецкая Н.М., Клинов Д.В., Бобров М.Ю., Шибанова Е.Д., Акимов М.Г., Фомина-Агеева Е.В., Зинченко Г.Н., Баирамашвили Д.И., Мирошников А.И. (2009). Нанокомплексы рекомбинантных белков с полисиаловой кислотой. Получение, свойства и биологическая активность. Биоорг. хим. 35 (3), 350–356 ID:197
  4. Таран С.А., Верёвкина К.Н., Феофанов С.А., Мирошников А.И. (2009). Ферментативное трансгликозилирование природных и модифицированных нуклеозидов иммобилизованными термостабильными нуклеозидфосфорилазами из Geobacillus stearothermophilus. Биоорг. хим. 35 (6), 822–829 ID:198
  5. Kayushin A., Korosteleva M., Miroshnikov A. (2009). Large-scale solid-phase preparation of 3'-unprotected trinucleotide phosphotriesters--precursors for synthesis of trinucleotide phosphoramidites. Nucleosides Nucleotides Nucleic Acids 19 (10-12), 1967–76 [+]

    The approach to large-scale solid-phase synthesis of 3'-unprotected trinucleotide phosphotriesters has been developed. The trinucleotides have been synthesized in 5 g scale by phosphotriester approach using CPG with pore size 70A. Total yield of target products was 75-90%. The molar extinctions of trinucleotides at various wave-lengths were calculated; the experimental UV-spectra of trinucleotides show a good agreement with theoretical ones. The trinucleotides synthesized were used for synthesis of trinucleotide phosphoramidites - synthons for generation of DNA/peptide libraries.

    ID:205
  6. Romanov V.P., Bezuglov V.V., Bobrov M.I.u., Kostromina T.I., Feofanov S.A., Miroshnikov A.I. (2009). [Isolation of expressed in E. coli human interferon beta1b (Ser17) by ion-exchange chromatography]. Bioorg. Khim. 37 (3), 327–33 [+]

    A method for isolation of interferon beta1b (Serl7) from inclusion bodies, comprising the steps of solution and reduction of protein from the inclusion bodies, refolding, chromatography on DEAE-Sepharose, chromatography on SP-Sepharose, concentrating, desalting and addition of stabilizers. The solution of reduced protein was diluted with pH 8.0 buffer of 50 mM Tris-HCl, 25 microM CuCl2 and 0.5% Twin 20 for refolding. We used gradient of pH (from 9.3 upto 11.3) for elution of interferon-beta from cation-exchange column. We concentrated of eluate and then desalted on the Sephadex G-50 column with 1 mM NaOH. Then the protein solution was neutralized with mannitol and Na-phosphate. Obtained preparation of interferon-beta was pure by gel-electrophoresis and by HPLC analysis, and had practically indentical level of antiproliferative activity with well-known preparation of Betaferone. Thus we show the possibility of isolation and obtaining of pure and active interferone-beta by ion-exchange chromatography in the presence of non-ion detergent Twin 20. We believe this method for interferon betalb preparation is perspective for scaling and using in the develop of industrial technology for production of this preparation.

    ID:581
  7. Скоблов А.Ю., Микулинская Г.В., Таран С.А., Мирошников А.И., Феофанов С.А., Скоблов Ю.С. (2009). Субстратная специфичность дезоксирибонуклеозидмонофосфаткиназы фага Т5 и ее использование для синтеза [α-32P]d/rNTP. Bioorg. Khim. 35 (6), 816–821 ID:2097
  8. Таран С.А., Верёвкина К.Н., Есикова Т.З., Феофанов С.А., Мирошников А.И. (2008). Синтез 2-хлор-2’-дезоксиаденозина микробиологическим трансгликозилированием с использованием рекомбинантного штамма Escherichia coli. Прикладная биохимия и микробиология 44 (2), 181–186 ID:2095
  9. Микулинская Г.В., Зимин А.А., Феофанов С.А., Мирошников А.И. (2007). Новая широкоспецифичная дезоксирибонуклеозидмонофосфаткиназа, кодируемая геном 52 бактериофага øС31. ДАН 412, 15–17 ID:2094
  10. Yagodkin A., Azhayev A., Roivainen J., Antopolsky M., Kayushin A., Korosteleva M., Miroshnikov A., Randolph J., Mackie H. (2007). Improved synthesis of trinucleotide phosphoramidites and generation of randomized oligonucleotide libraries. Nucleosides Nucleotides Nucleic Acids 26 (5), 473–97 [+]

    A new method to produce a set of 20 high quality trinucleotide phosphoramidites on a 5-10 g scale each was developed. The procedure starts with condensation reactions of P-components with N-acyl nucleosides, bearing the 3 '-hydroxyl function protected with 2-azidomethylbenzoyl, to give fully protected dinucleoside phosphates 13. Upon cleavage of dimethoxytrityl group from 13, dinucleoside phosphates 16 are initially transformed into trinucleoside diphosphates 19 and then the 2-azidomethylbenzoyl is selectively removed under neutral conditions to generate trinucleoside diphosphates 5 in excellent yield. Subsequent 3 '-phosphitylation affords target trinucleotide phosphoramidites 7. When mutagenic oligonucleotides are synthesized employing mixtures of building blocks 7 as well as following the new synthetic protocol, representative oligonucleotide libraries are generated in good yields.

    ID:199
  11. Roivainen J., Elizarova T., Lapinjoki S., Mikhailopulo I.A., Esipov R.S., Miroshnikov A.I. (2007). An enzymatic transglycosylation of purine bases. Nucleosides Nucleotides Nucleic Acids 26 (8-9), 905–9 [+]

    An enzymatic transglycosylation of purine heterocyclic bases employing readily available natural nucleosides or sugar-modified nucleosides as donors of the pentofuranose fragment and recombinant nucleoside phosphorylases as biocatalysts has been investigated. An efficient enzymatic method is suggested for the synthesis of purine nucleosides containing diverse substituents at the C6 and C2 carbon atoms. The glycosylation of N(6)-benzoyladenine and N(2)-acetylguanine and its O(6)-derivatives is not accompanied by deacylation of bases.

    ID:200
  12. Chuvikovsky D.V., Esipov R.S., Skoblov Y.S., Chupova L.A., Muravyova T.I., Miroshnikov A.I., Lapinjoki S., Mikhailopulo I.A. (2006). Ribokinase from E. coli: expression, purification, and substrate specificity. Bioorg. Med. Chem. 14 (18), 6327–32 [+]

    Ribokinase (RK) was expressed in the Escherichia coli ER2566 cells harboring the constructed expression plasmid encompassing the rbsK gene, encoding ribokinase. The recombinant enzyme was purified from sonicated cells by double chromatography to afford a preparation that was ca. 90% pure and had specific activity of 75 micromol/min mg protein. Catalytic activity of RK: (i) is strongly dependent on the presence of monovalent cations (potassium>>>ammonium>cesium), and (ii) is cooperatively enhanced by divalent magnesium and manganese ions. Besides D-ribose and 2-deoxy-D-ribose, RK was found to catalyze the 5-O-phosphorylation of D-arabinose, D-xylose, and D-fructose in the presence of ATP, and potassium and magnesium ions; L-ribose and L-arabinose are not substrates for the recombinant enzyme. A new radiochemical method for monitoring the formation of D-pentofuranose-5-[32P]phosphates in the presence of [gamma-32P]ATP and RK is reported.

    ID:201
  13. Mikoulinskaia G.V., Zimin A.A., Feofanov S.A., Miroshnikov A.I. (2004). Identification, cloning, and expression of bacteriophage T5 dnk gene encoding a broad specificity deoxyribonucleoside monophosphate kinase (EC 2.7.4.13). Protein Expr. Purif. 33 (2), 166–75 [+]

    The nucleotide sequence corresponding to 13-19.5% of the bacteriophage T5 genome in early region C was determined (GenBank AY 140897). One of the five major single-stranded interruptions (nicks) of bacteriophage T5 DNA was identified at 18.5%. The sequenced region was annotated and the putative functions of some open reading frames were proposed by comparison with databases. The dnk gene, encoding a deoxyribonucleoside monophosphate kinase, was identified using a previously defined N-terminal amino acid sequence. The gene was cloned and expressed in Escherichia coli, the enzyme was purified to homogeneity with high yield using two alternative methods, and the recombinant deoxyribonucleoside monophosphate kinase was found to have the same activity and specificity as the native enzyme.

    ID:2092
  14. Константинова И.Д., Леонтьева Н.А., Галегов Г.А., Рыжова О.И., Чувиковский Д.В., Антонов К.А., Есипов Р.С., Таран С.А., Верёвкина К.Н., Феофанов С.А., Мирошников А.И. (2004). Биотехнологический способ получения рибавирина. Действие рибавирина и некоторых его комбинаций на репродукцию вируса осповакцины. Bioorg. Khim. 30 (6), 613–620 ID:2093
  15. Mikoulinskaia G.V., Gubanov S.I., Zimin A.A., Kolesnikov I.V., Feofanov S.A., Miroshnikov A.I. (2003). Purification and characterization of the deoxynucleoside monophosphate kinase of bacteriophage T5. Protein Expr. Purif. 27 (2), 195–201 [+]

    Deoxynucleoside monophosphate kinase (dNMP kinase) of bacteriophage T5 (EC 2.7.4.13) was purified to apparent homogeneity from phage-infected Escherichia coli cells. Electrophoresis in sodium dodecyl sulfate-polyacrylamide gel showed that the enzyme has a molecular mass of about 29 kDa. The molecular mass of dNMP kinase estimated by analytical equilibrium ultracentrifugation turned out to be 29.14 +/- 3.03 kDa. These data suggest that the enzyme exists in solution as a monomer. The isoelectric point of dNMP kinase was found to be 4.2. The N-terminal amino acid sequence, comprising 21 amino acids, was determined to be VLVGLHGEAGSGKDGVAKLII. A comparison of this amino acid sequence and those of known enzymes with a similar function suggests the presence of a nucleotide-binding site in the sequenced region.

    ID:202
  16. Антонов К.В., Есипов Р.С., Гуревич А.И., Чувиковский Д.В., Микулинская Г.В., Феофанов С.А., Мирошников А.И. (2003). Химический и химико-ферментативный синтез α-тиотрифосфатов нуклеозидов. Bioorg. Khim. 29 (6), 616–622 ID:2091
  17. Esipov R.S., Gurevich A.I., Chuvikovsky D.V., Chupova L.A., Muravyova T.I., Miroshnikov A.I. (2002). Overexpression of Escherichia coli genes encoding nucleoside phosphorylases in the pET/Bl21(DE3) system yields active recombinant enzymes. Protein Expr. Purif. 24 (1), 56–60 [+]

    The Escherichia coli genes encoding purine nucleoside phosphorylase, uridine phosphorylase, and thymidine phosphorylase were cloned into pET plasmids to generate highly effective E. coli BL21(DE3) strains producing each of these enzymes. Optimum conditions for biosynthesis of each enzyme as a soluble protein with intact biological activity were found. The crude preparations are approximately 80% pure and can be used immediately for enzymatic transglycosylation. The enzyme preparations were purified to homogeneity by two steps including fractional precipitation with ammonium sulfate and subsequent chromatography on Sephadex G-100 and DEAE-Sephacel.

    ID:203
  18. Sergeev N.V., Gloukhova N.S., Nazimov I.V., Gulyaev V.A., Shvets S.V., Donetsky I.A., Miroshnikov A.I. (2001). Monitoring of recombinant human insulin production by narrow-bore reversed-phase high-performance liquid chromatography, high-performance capillary electrophoresis and matrix-assisted laser desorption ionisation time-of-flight mass spectrometry. J Chromatogr A 907 (1-2), 131–44 [+]

    An analytical scheme for monitoring recombinant human insulin (rhI) production is suggested. The scheme includes high-performance separation micro-techniques (narrow-bore RP-HPLC, HPCE) based on different separation mechanisms and matrix-assisted laser desorption ionisation time-of-flight MS, and allows one to obtain unambiguous information about purity and primary structure of all intermediates of the rhI production. The use of this scheme at all production steps provided optimisation of certain technological parameters [conditions for a fusion protein (FP) refolding, temperature and duration of the FP cleavage with trypsin, conditions for carboxypeptidase B digestion of di-ArgB31-B32-insulin] and achievement of a high purity of the end-product. The proposed scheme may be used for solving various problems in monitoring production of other recombinant proteins.

    ID:204
  19. Миргородская О.А., Козьмин Ю.П., Титов М.И., Савельева Н.М., Кернер Р., Сонксен К., Ройпсторфф П., Мирошников А.И. (2000). Использование MALDI-MS для количественного анализа пептидов и белков. Биоорг. хим. 26 (9), 662–671 ID:206
  20. Martynov V.I., Kostina M.B., Feigina M.I.u., Miroshnikov A.I. (1983). Limited proteolysis studies on molecular organization of bovine rhodopsin in the photoreceptor membrane. Bioorg Khim. 9, 734–745 [+]

    By limited proteolysis of rhodopsin in disk-membranes of native (extradisc-out) and inverted (intradisc-out) orientation, it was shown for the first time that the protein consists of seven transmembrane domains traversing the membrane. Polypeptide fragments exposed to the cytoplasmic and intradiscal space of a photoreceptor cell, and the sequences embedded into phospholipid matrix have been determined.

    ID:65
  21. Shemyakin M.M., Ovchinnikov Yu.A., Kiryushkin A.A., Vinogradova E.I., Miroshnikov A.I., Alakhov Yu.B., Lipkin V.M., Shvetsov Yu.B., Wulfson N.S., Rosinov B.V., Bochkarev V.N., Burikov V.M. (1966). Mass spectrometric determination of the amino-acid sequence of peptides. Nature 211 (5047), 361–6 [+]

    New method of determination polypeptide amino acids sequenses with mass-spectoscopy of acyl ethers. This method base on peptide fragmentation with localizing positive charge on N-acyl fragments.

    ID:167