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 Персоналии
Eugene S. Vysotski
Head of Photobiology Laboratory
Institute of Biophysics of the Russian Academy of Sciences, Siberian Branch
Akademgorodok 50, Bldg. 50, Krasnoyarsk 660036, Russia
Educational background:
Krasnoyarsk State University, Krasnoyarsk, Russia M.S. in Biophysics 1975
Institute of Biophysics of the Russian Academy of Sciences
Siberian Branch, Krasnoyarsk, Russia PhD in Biophysics 1987
Graduate advisor: Josef I. Gitelson, academician, Institute of Biophysics, RAS, SB, Russia
Thesis title: The studies on the nature of an aldehyde factor – the endogenous substrate of the luminous bacteria luciferase
Professional appointments:
Head of Photobiology Lab
Institute of Biophysics RAS, SB, Krasnoyarsk, Russia 1991-present
Senior Research Scientist, Photobiology Lab
Institute of Biophysics RAS, SB, Krasnoyarsk, Russia 1989-1991
Research Scientist, Photobiology Lab
Institute of Biophysics RAS, SB, Krasnoyarsk, Russia 1987-1989
Junior Research Scientist, Photobiology Lab
Institute of Biophysics RAS, SB, Krasnoyarsk, Russia 1982-1987
Assistant, Photobiology Lab
Institute of Biophysics RAS, SB, Krasnoyarsk, Russia 1975-1982
Visiting Scholar (various months)
Friday Harbor Laboratories, University of Washington 1994-2001
Visiting Scholar (various months)
University of Georgia, Dept. of Biochemistry and Molecular Biology 1998-2006
Visiting Scholar (several months)
Wageningen University, The Netherlands 2007
Associate Research Scientist
University of Georgia, Dept. of Biochemistry and Molecular Biology 2000-2002
Research Interests:
Publications:
Publications in the last five years:
Larionova M.D., Markova S.V., and Vysotski E.S. (2018) Bioluminescent and structural features of native folded Gaussia luciferase. J. Photochem. Photobiol. B. 183, 309-317.
Eremeeva E.V. and Vysotski E.S. (2018) Exploring bioluminescence function of the Ca2+-regulated photoproteins with site-directed mutagenesis. Photochem. Photobiol., doi: 10.1111/php.12945.
Larionova M.D., Markova S.V. and Vysotski E.S. (2017) Tyr72 and Tyr80 are involved in the formation of an active site of a luciferase of copepod Metridia longa. Photochem. Photobiol. 93, 503-510.
Markova S.V, Malikova N.P., Vysotski E.S., Frank L.A. and Gitelson I.I. (2017) Bioluminescent monitoring enables observation of intracellular events in real time without cell and tissue destruction. Biophysics 62, No. 3, 503-507.
Eremeeva E.V., Bartsev S.I., van Berkel W.J.H. and Vysotski E.S. (2017) Unanimous model for describing the fast bioluminescence kinetics of Ca2+-regulated photoproteins of different organisms. Photochem. Photobiol. 93, 495-502.
Larionova M.D., Markova S.V. and Vysotski E.S. (2017) The novel extremely psychrophilic luciferase from Metridia longa: Properties of a high-purity protein produced in insect cells. Biochem. Biophys. Res. Commun. 483, 772-778.
Vysotski E.S. (2017) Bioluminescence: Basic and Applied Research: Introduction to the Special Issue on Bioluminescence. Photochem. Photobiol. 93, 387-388.
Eremeeva E.V. and Vysotski E.S. (2017) Bioluminescent and biochemical properties of Cys-free Ca2+-regulated photoproteins obelin and aequorin. J. Photochem. Photobiol. B. 174, 97-105.
Markova S.V., Larionova M.D., Gorbunova D.A. and Vysotski E.S. (2017) The disulfide-rich Metridia luciferase refolded from E. coli inclusion bodies reveals the properties of a native folded enzyme produced in insect cells. J. Photochem. Photobiol. B. 175, 51-57.
Eremeeva E.V., van Berkel W.J. and Vysotski E.S. (2016) Transient-state kinetic analysis of complex formation between photoprotein clytin and GFP from jellyfish Clytia gregaria. FEBS Lett 590, 307-316.
Burakova L.P., Natashin P.V., Malikova N.P., Niu F., Pu M., Vysotski E.S. and Liu Z.J. (2016) All Ca2+-binding loops of light-sensitive ctenophore photoprotein berovin bind magnesium ions: The spatial structure of Mg2+-loaded apo-berovin. J. Photochem. Photobiol. B. 154, 57-66.
Burakova L.P., Stepanyuk G.A., Eremeeva E.V. and Vysotski E.S. (2016) Role of certain amino acid residues of coelenterazine-binding cavity in bioluminescence of light-sensitive Ca2+-regulated photoprotein berovin. Photochem. Photobiol. Sci. 15, 691-704.
Burakova L.P., Natashin P.V., Markova S.V., Eremeeva E.V., Malikova N.P., Cheng C., Liu Z.J. and Vysotski E.S. (2016) Mitrocomin from the jellyfish Mitrocoma cellularia with deleted C-terminal tyrosine reveals a higher bioluminescence activity compared to wild type photoprotein. J. Photochem. Photobiol. B. 162, 286-297.
Markova S.V. and Vysotski E.S. (2015) Coelenterazine-dependent luciferases. Biochemistry (Mosc) 80, 714-732.
Markova, S.V., Larionova, M.D., Burakova, L.P. and Vysotski, E.S. (2015) The smallest natural high-active luciferase: cloning and characterization of novel 16.5-kDa luciferase from copepod Metridia longa. Biochem .Biophys. Res. Commun. 457, 77-82.
Malikova, N.P., Borgdorff, A.J. and Vysotski, E.S. (2015) Semisynthetic photoprotein reporters for tracking fast Ca2+ transients. Photochem. Photobiol. Sci. 14, 2213-2224.
Natashin, P.V., Ding, W., Eremeeva, E.V., Markova, S.V., Lee, J., Vysotski, E.S. and Liu, Z.J. (2014) Structures of the Ca2+-regulated photoprotein obelin Y138F mutant before and after bioluminescence support the catalytic function of a water molecule in the reaction. Acta Crystallogr. D Biol. Crystallogr. D70, 720-732.
Natashin, P.V., Markova, S.V., Lee, J., Vysotski, E.S. and Liu, Z.J. (2014) Crystal structures of the F88Y obelin mutant before and after bioluminescence provide molecular insight into spectral tuning among hydromedusan photoproteins. FEBS J. 281, 1432-1445.
Gealageas, R., Malikova, N.P., Picaud, S., Borgdorff, A.J., Burakova, L.P., Brulet, P., Vysotski, E.S. and Dodd, R.H. (2014) Bioluminescent properties of obelin and aequorin with novel coelenterazine analogues. Anal. Bioanal. Chem. 406, 2695-2707.
Malikova, N.P., Burakova, L.P., Markova, S.V. and Vysotski, E.S. (2014) Characterization of hydromedusan Ca2+-regulated photoproteins as a tool for measurement of [Ca2+]. Anal. Bioanal. Chem. 406, 5715-5726.
Selected peer-reviewed publications:
Illarionov, B.A., Bondar, V.S., Illarionova, V.A., and Vysotski, E.S. (1995) Sequence of the cDNA encoding the Ca2+-activated photoprotein obelin from the hydroid polyp Obelia longissima. Gene 153, 273-274.
Vysotski, E.S., Liu, Z.J., Rose, J., Wang, B.C., and Lee, J. (1999) Preparation and preliminary study of crystals of the recombinant calcium-regulated photoprotein obelin from the bioluminescent hydroid Obelia longissima. Acta Crystallogr. D Biol. Crystallogr. D55, 1965-1966.
Liu, Z.J., Vysotski, E.S., Chen, C.J., Rose, J.P., Lee J., and Wang, B.C. (2000) Structure of the Ca2+-regulated photoprotein obelin at 1.7 A resolution determined directly from its sulfur substructure. Protein Sci. 9, 2085-2093.
Deng, L., Vysotski, E.S., Liu, Z.J., Markova, S.V., Malikova, N.P., Lee, J., Rose, J., and Wang, B.C. (2001) Structural basis for the emission of violet bioluminescence from a W92F obelin mutant. FEBS Lett. 506, 281-285.
Vysotski, E.S., Liu, Z.J., Rose, J., Wang, B.C., and Lee, J. (2001) Preparation and X-ray crystallographic analysis of recombinant obelin crystals diffracting to beyond 1.1 A. Acta Crystallogr. D Biol. Crystallogr. D57, 1919-1921.
Markova, S.V., Vysotski, E.S., Blinks, J.R., Burakova, L.P., Wang, B.C., and Lee J. (2002) Obelin from the bioluminescent marine hydroid Obelia geniculata: cloning, expression, and comparison of some properties with those of other Ca2+-regulated photoproteins. Biochemistry 41, 2227-2236.
Vysotski, E.S., Liu, Z.J., Markova, S.V., Blinks, J.R., Deng, L., Frank, L.A., Herko, M., Malikova, N.P., Rose, J.P., Wang, B.C., and Lee, J. (2003) Violet bioluminescence and fast kinetics from W92F obelin: Structure-based proposals for the bioluminescence triggering and the identification of the emitting species. Biochemistry 42, 6013-6024.
Malikova, N.P., Stepanyuk, G.A., Frank, L.A., Markova, S.V., Vysotski, E.S. and Lee J. (2003) Spectral tuning of obelin bioluminescence by mutations of Trp92. FEBS Lett. 554: 184-188.
Liu, Z.J., Vysotski, E.S., Deng, L., Lee, J., Rose, J., and Wang, B.C. (2003) Atomic resolution structure of obelin: soaking with calcium enhances electron density of the second oxygen atom substituted at the C2-position of coelenterazine. Biochem. Biophys. Res. Commun. 311, 433-439.
Deng, L., Markova, S.V., Vysotski, E.S., Liu, Z.J., Lee, J., Rose, J., and Wang, B.C. (2004) Preparation and X-ray crystallographic analysis of the Ca2+-discharged photoprotein obelin. Acta Crystallogr. D Biol. Crystallogr. D60, 512-514.
Deng, L., Markova, S.V., Vysotski, E.S., Liu, Z.J., Lee, J., Rose, J., and Wang, B.C. (2004) Crystal structure of a Ca2+-discharged photoprotein: Implications for mechanisms of the calcium trigger and bioluminescence. J. Biol. Chem. 279, 33647-33652.
Markova, S.V., Golz, S., Frank, L.A., Kalthof, B., and Vysotski, E.S. (2004) Cloning and expression of cDNA for a luciferase from the marine copepod Metridia longa. A novel secreted bioluminescent reporter enzyme. J. Biol. Chem. 279, 3212-3217.
Vysotski, E.S. and Lee, J. (2004) Ca2+-regulated photoproteins: structural insight into the bioluminescence mechanism. Acc. Chem. Res. 37, 405-415.
Stepanyuk, G.A., Golz, S., Markova, S.V., Frank, L.A., Lee, J., and Vysotski, E.S. (2005) Interchange of aequorin and obelin bioluminescence color is determined by substitution of one active site residue of each photoprotein. FEBS Lett. 579, 1008-1014.
Deng, L., Vysotski, E.S., Markova, S.V., Liu, Z.-J., Lee, J., Rose, J., and Wang, B.C. (2005) All three Ca2+-binding loops of photoproteins bind calcium ions: the crystal structures of calcium-loaded apo-aequorin and apo-obelin. Protein Sci. 14, 663-675.
Liu, Z.J., Stepanyuk, G.A., Vysotski, E.S., Lee, J., Markova, S.V., Malikova, N.P., and Wang, B.C. (2006) Crystal structure of obelin after Ca2+-triggered bioluminescence suggests neutral coelenteramide as the primary excited state. Proc. Natl. Acad. Sci. USA 103, 2570-2575.
Vysotski, E.S., Markova, S.V., Frank, L.A. (2006) Calcium-regulated photoproteins of marine coelenterates. Molecular Biology 40, 355-367.
Titushin, M.S., Markova, S.V., Frank, L.A., Malikova, N.P., Stepanyuk, G.A., Lee, J., and Vysotski, E.S. (2008) Coelenterazine-binding protein of Renilla muelleri: cDNA cloning, overexpression, and characterization as a substrate of luciferase. Photochem. Photobiol. Sci. 7, 189-196.
Stepanyuk, G.A., Liu, Z.J., Markova, S.V., Frank, L.A., Lee, J., Vysotski, E.S., and Wang, B.C. (2008) Crystal structure of coelenterazine-binding protein from Renilla muelleri at 1.7 A: Why it is not a calcium-regulated photoprotein. Photochem Photobiol Sci. 7, 442-447.
Frank, L.A., Borisova, V.V., Markova, S.V., Malikova, N.P., Stepanyuk, G.A., and Vysotski, E.S. (2008) Violet and greenish photoprotein obelin mutants for reporter applications in dual-color assay. Anal. Bioanal. Chem. 391, 2891-2896.
Borisova, V.V., Frank, L.A., Markova, S.V., Burakova, L.P., and Vysotski, E.S. (2008) Recombinant Metridia luciferase isoforms: expression, refolding and applicability for in vitro assay. Photochem. Photobiol. Sci. 7, 1025-1031.
Stepanyuk, G.A., Liu, Z.J., Vysotski, E.S., Lee, J., Rose, J.P., and Wang, B.C. (2009) Structure based mechanism of the Ca2+-induced release of coelenterazine from the Renilla binding protein. Proteins 74, 583-593.
Eremeeva, E.V., Markova, S.V., Westphal, A.H., Visser, A.J., van Berkel, W.J., and Vysotski, E.S. (2009) The intrinsic fluorescence of apo-obelin and apo-aequorin and use of its quenching to characterize coelenterazine binding. FEBS Lett. 583, 1939-1944.
van Oort, B., Eremeeva, E.V., Koehorst, R.B., Laptenok, S.P., van Amerongen, H., van Berkel, W.J., Malikova, N.P., Markova, S.V., Vysotski, E.S., Visser, A.J., and Lee, J. (2009) Picosecond fluorescence relaxation spectroscopy of the calcium-discharged photoproteins aequorin and obelin. Biochemistry 48, 10486-10491.
Markova, S.V., Burakova, L.P., Frank, L.A., Golz, S., Korostileva, K.A., and Vysotski, E.S. (2010) Green-fluorescent protein from the bioluminescent jellyfish Clytia gregaria: cDNA cloning, expression, and characterization of novel recombinant protein. Photochem. Photobiol. Sci. 9, 757-765.
Stepanyuk, G.A., Unch, J., Malikova, N.P., Markova, S.V., Lee, J., and Vysotski, E.S. (2010) Coelenterazine-v ligated to Ca2+-triggered coelenterazine-binding protein is a stable and efficient substrate of the red-shifted mutant of Renilla muelleri luciferase. Anal. Bioanal. Chem. 398, 1809-1817.
Titushin, M.S., Feng, Y., Stepanyuk, G.A., Li, Y., Markova, S.V., Golz, S., Wang, B.C., Lee, J., Wang, J., Vysotski, E.S., and Liu, Z.J. (2010) NMR-derived topology of a GFP-photoprotein energy transfer complex. J. Biol. Chem. 285, 40891-40900.
Malikova, N.P., Visser, N.V., van Hoek, A., Skakun, V.V., Vysotski, E.S., Lee, J., and Visser, A.J. (2011) Green-fluorescent protein from the bioluminescent jellyfish Clytia gregaria is an obligate dimer and does not form a stable complex with the Ca2+-discharged photoprotein clytin. Biochemistry 50, 4232-4241.
Markova, S.V., Burakova, L.P., and Vysotski, E.S. (2012) High-active truncated luciferase of copepod Metridia longa. Biochem. Biophys. Res. Commun. 417, 98-103.
Markova, S.V., Burakova, L.P., Golz, S., Malikova, N.P., Frank, L.A., and Vysotski, E.S. (2012) The light-sensitive photoprotein berovin from the bioluminescent ctenophore Beroe abyssicola: a novel type of Ca2+-regulated photoprotein. FEBS J. 279, 856-870.
Titushin, M.S., Feng, Y., Lee, J., Vysotski, E.S., and Liu, Z.J. (2011) Protein-protein complexation in bioluminescence. Protein & Cell 2, 957-972.
Eremeeva, E.V., Vysotski, E.S., Westphal, A.H., van Mierlo, C.P., van Berkel, W.J. (2012) Ligand binding and conformational states of the photoprotein obelin. FEBS Lett. 586, 4173-4179.
Eremeeva, E.V., Natashin, P.V., Song, L., Zhou, Y., van Berkel, W.J., Liu, Z.J., and Vysotski, E.S. (2013) Oxygen activation of apo-obelin-coelenterazine complex. ChemBioChem 14, 739-745.
Eremeeva, E.V., Markova, S.V., Frank, L.A., Visser, A.J., van Berkel, W.J., and Vysotski, E.S. (2013) Bioluminescent and spectroscopic properties of His-Trp-Tyr triad mutants of obelin and aequorin. Photochem. Photobiol. Sci. 12, 1016-1024.
Eremeeva, E.V., Markova, S.V., and Vysotski, E.S. (2013) Highly active BRET-reporter based on yellow mutant of Renilla muelleri luciferase. Dokl. Biochem. Biophys. 450, 147-150.
Stepanyuk, G.A., Liu, Z.J., Burakova, L.P., Lee, J., Rose, J., Vysotski, E.S., and Wang, B.C. (2013) Spatial structure of the novel light-sensitive photoprotein berovin from the ctenophore Beroe abyssicola in the Ca2+-loaded apoprotein conformation state. Biochim. Biophys. Acta 1834, 2139-2146.
Eremeeva, E.V., Markova, S.V., van Berkel, W.J., and Vysotski, E.S. (2013) Role of key residues of obelin in coelenterazine binding and conversion into 2-hydroperoxy adduct. J. Photochem. Photobiol. B 127, 133-139.
Supervisor experience:
Vladimir S. Bondar PhD thesis title: Ca2+-regulated photoprotein obelin from the hydroid Obelia longissima
Ludmila A. Frank PhD thesis title: The recombinant photoprotein obelin from the hydroid Obelia longissima: isolation and application in immunoassay
Victoria A. Illarionova PhD thesis title: Ca2+-regulated photoprotein obelin from the hydroid Obelia longissima: investigation of structural and functional relations using site-directed mutagenesis
alina A. Stepanyuk PhD thesis title: Structure-function features of calcium-regulated coelenterazine-binding proteins
Maxim S. Titushin PhD thesis title: Protein-protein interactions in bioluminescence systems of coelenterates Renilla muelleri and Clytia gregaria
Elena V. Eremeeva PhD thesis title: Active photoprotein complex formation using obelin and aequorin as well as their mutants as an example
and PhD thesis title: Molecular mechanism of active photoprotein complex formation (Wageningen University, The Netherlands)
Pavel V. Natashin PhD thesis title: Function of some amino acid residues in bioluminescence of Ca2+-regulated photoproteins
Ludmila P. Burakova PhD thesis title: Photosensitive photoprotein berovin of ctenophore Beroe abyssicola: cloning and properties of recombinant protein
Marina D. Larionova PhD thesis title: The novel isoforms of luciferase from copepod Metridia longa: properties and application
Professional affiliations:
Associate Editor, Photochemistry and Photobiology,
Journal of American Society of Photobiology 2013-present
Expert of the Russian Foundation for Basic Research 2003-present
Member of the Expert Board on biological and medical sciences of the Russian Foundation for Basic Research 2007-2012
Recent patents:
Golz, S., Markova, S., Burakova, L., Frank, L. & Vysotski, E. Isolated photoprotein mtclytin, and use thereof. WO 2005035559, April 21, 2005.
Golz, S., Markova, S., Burakova, L., Frank, L. & Vysotski, E. Isolated berovin photoprotein and use thereof. WO 2005021591, March 10, 2005.
Golz, S., Markova, S., Burakova, L., Frank, L. & Vysotski, E. Isolated photoprotein bolinopsin, and the use thereof. WO 2005000885, January 6, 2005.
Golz, S., Markova, S., Burakova, L., Frank, L. & Vysotski, E. Isolated fluorescent protein cgfp from Clytia gregarium and the use thereof. WO 2004052926, June 24, 2004.
Golz, S., Kalthof, B., Markova, S., Frank, L. & Vysotski, E. Isolated luciferases lu164, luAl and lu22, and the use of the same. WO 0242469, May 30, 2002.
Borisova, V.V., Frank, L.A., Markova, S.V., Vysotski, E.S. Method of simultaneous determination of two analytes by bioluminescence molecular microanalysis. Patent RF No. 2373540 (20.11.2009).
Malikova, N.P., Vysotski, E.S., Markova, S.V., Stepanyuk, G.A. Mutant photoprotein for simultaneous measurement of intracellular calcium in different cell compartments. Patent RF No. 2402566 (27.10.2010).
Markova, S.V., Vysotski, E.S., Natashin, P.V. Composition of the complementing fragments of photoprotein from Hydrozoa and method of its application for determination of molecular interactions. Patent RF No. 2402566 (20.05.2011).
Markova, S.V., Burakova, L.P., Vysotski, E.S. Truncated mutant luciferase from Metridia longa for application as a bioluminescence reporter in mammalian cells. Patent RF No. 2495929 (20.10.2013).
Frank, L.A., Kudryavtsev, A.N., Krasitskaya, V.V., Vysotski, E.S. Method of simultaneous determination of two analytes by bioluminescence molecular microanalysis. Patent RF No. 2011154712 (27.10.2013).
Research support in the last five years:
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