Prof. Dr hab. Borys Kierdaszuk

Division of Biophysics
Institute of Experimental Physics
University of Warsaw
Zwirki i Wigury 93, 02-089 Warsaw, Poland

Phone: +48 22 5540784

E-mail: borys@biogeo.uw.edu.pl

Research interests

Mechanism of action of biological molecules (e.g. enzymes, chloroplasts) and their constituents (e.g. tryptophan, tyrosine, chlorophylls), with the aid of emission spectroscopy (fluorescence, phosphorescence) and enzyme kinetics, including time-resolved techniques, as follows:

  1. Emission (fluorescence, phosphorescence) spectroscopy applied in biophysical studies of bio-macromolecular systems and their constituents, to gain knowledge on the mechanism of recognition and kinetics of protein-ligand binding, e.g. photosynthetic protein-chromofor, enzyme-substrate and enzyme-inhibitor complexes;
  2. Identification and characterization of reaction transition states;
  3. Effect of ligand binding on its conformation and electronic structure;
  4. Confrontation of solution studies with crystallographic data.

To better understand the molecular mechanisms of key biological processes, e.g. catalysis, photosynthesis etc., towards development of sensitive and selective methods of detection in vitro and in vivo.

Professional experience

  1. Experience in different kinds of instrumentation for absorption and emission measurements, including time-domain single-photon counting for fluorescence lifetime and anisotropy determination
  2. Characterization and identification of molecular structures using spectroscopic (1H, 13C and 15N NMR, CD, IR, and electronic absorption and emission) chromatographic methods (TLC, HPLC, FPLC, column chromatography)
  3. Photochemistry and photobiophysics of biopolymers and their constituents
  4. Inter- and intramolecular hydrogen bonding interactions
  5. Intra- and Intermolecular hydrogen exchange and various phototropic equilibria including tautomerism of nucleosides and their analogues, and proton association-dissociation in biomacromolecules
  6. Physical basis of heterogeneity of protein fluorescence
  7. Protein-ligand interactions, and their effect on protein dynamics and fluorescence quenching
  8. Purification and characterization of proteins using chromatographic, hydrodynamic, electrophoretic and spectroscopic techniques as well as measurements of protein (enzyme) activity
  9. High precision sample preparation in micro and macro scales
  10. Separation and purification of organic compounds using chromatographic techniques (TLC, HPLC, FPLC, column chromatography), hydrodynamic methods, extraction, sublimation and crystallization
  11. Analysis of experimental data using statistical software
1979 - 1981 Department of Biophysics, Institute of Experimental Physics, University of Warsaw, Warsaw, Poland; Ph.D. student
1981 - 1986 Department of Biophysics, Institute of Experimental Physics, University of Warsaw, Warsaw, Poland; Research Assistant
1986 - 1988 Karolinska Institute, Stockholm, Sweden; Postdoctoral Research Associate (Fellowship of Swedish Research Council)
1988 - 1994 Department of Biophysics, University of Warsaw, Warsaw, Poland; Adjunct
1992 - Department of Medical Biophysics, Karolinska Institute, Stockholm, Sweden; Postdoctoral Research Associate (EMBO Fellowship)
1994 - 1995 Center for Fluorescence Spectroscopy, Department of Biological Chemistry, University of Maryland at Baltimore, Baltimore, Maryland, USA; Postdoctoral Research Associate (Fulbright Fellow)
1995 - 1999 Department of Biophysics, University of Warsaw, Warsaw, Poland; Adjunct
1997, 2000 Department of Veterinary Chemistry, Biomedical Center, Uppsala, Sweden; Research Ass. Professor
1997 - Department of Life Sciences and Chemistry, Roskilde University, Roskilde, Denmark; Research Ass. Professor
1999 - Department of Biophysics, Institute of Experimental Physics, University of Warsaw, Warsaw, Poland; Ass. Professor

Selected publications (1995-2003)

  1. Włodarczyk J. and Kierdaszuk B., Interpretation of fluorescence decays using a power-like model. Biophysical Journal, 85(1), in press (2003)
  2. Krawiec K., Kierdaszuk B. and Shugar D., Inorganic tripolyphosphate (PPPi) as a phosphate donor for human deoxyribonucleoside kinases. Biochemical and Biophysical Research Communications 301, 192-197 (2003)
  3. Poznański J., Kierdaszuk B. and Shugar D., Structural properties of the neutral and monoanioic forms of xanthosine, highly relevant to their substrate properties with various enzyme systems. Nucleosides, Nucleotides & Nucleic Acids, 22, 249-263 (2003)
  4. Krawiec K., Kierdaszuk B., Kalinichenko E. N., Rubinova E. B., Mikhailopulo I. A., Eriksson S., Munch-Petersen B. and Shugar D., Striking ability of adenosine-2'(3')-deoxy-3'(2')-triphosphates, and related analogues, to replace ATP as phosphate donor for all human, and Drosphila melanogaster, deoxyribonucleoside kinases. Nucleosides, Nucleotides & Nucleic Acids, 22, 153-173 (2003)
  5. Kierdaszuk B., Emission spectroscopy of complex formation between Escherichia coli purine nucleoside phosphorylase (PNP) and identified tautomeric species of formycin inhibitors resolves ambiguities found in crystallographic studies. Fluorescence Spectroscopy, Imaging and Probes 2, 277-296 (2002)
  6. Stoychev G., Kierdaszuk B. and Shugar D., Xanthosine and xanthine: Substrate properties with purine nucleoside phosphorylases, and relevance to other enzyme systems. European Journal of Biochemistry 269, 4048-4057 (2002)
  7. Stoychev G., Kierdaszuk B. and Shugar D., Interaction of E. coli purine nucleoside phosphorylase (PNP) with the cationic and zwitterionic forms of the fluorescent substrate N(7)-methylguanosine. Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology, 1544, 74-88 (2001)
  8. Kierdaszuk B., Modrak-Wójcik A., Wierzchowski J. and Shugar D., Formycin A and its N-methyl analogues, specific inhibitors of E. coli purine nucleoside phosphorylase (PNP): induced tautomeric shifts on binding to enzyme, and enzyme-ligand fluorescence resonance energy transfer. Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology 1476, 109-128 (2000)
  9. Stepanenko T., Lapinski L., Sobolewski A.L., Nowak M.J. and Kierdaszuk B., Photochemical syn-anti isomerisation reaction in 1-methyl-N4-hydroxycytosine. An experimental matrix isolation and theoretical density functional theory study. Journal of Physical Chemistry, 104, 9459-9466 (2000)
  10. Kierdaszuk B., Krawiec K., Kazimierczuk Z., Jacobsson U., Johansson N.G., Munch Petersen B., Eriksson S. and Shugar D., Substrate/inhibitor specificities of human deoxycytidine kinase (dCK) and thymidine kinase (TK1 and TK2) towards the sugar moiety of nucleosides, including O'-alkyl analogues. Nucleosides & Nucleotides 18, 1883-1903 (1999)
  11. Kierdaszuk B., Krawiec K., Kazimierczuk Z., Jacobsson U., Johansson N.G., Munch-Petersen B., Eriksson S. and Shugar D., Substrate/inhibitor specificities of human deoxycytidine kinase (dCK) and thymidine kinase (TK1 and TK2). Advances in Experimental Medicine and Biology 431, 623-627 (1998)
  12. Niedzwiecka-Kornas A., Kierdaszuk B., Stolarski R. and Shugar D. Tautomerism, acid-base properties and conformation of methylated analogues of the promutagenic N4-hydroxycytosine. Biophysical Chemistry 71, 87-98 (1998)
  13. Krawiec K., Kierdaszuk B., Kalinichenko E.N., Mikhailopulo I.A. and Shugar D., Unusual nucleoside triphosphate donors for nucleoside kinases: 3'-deoxyadenosine-2'-triphosphate and 2'-deoxyadenosine-3'-triphosphate. Acta Biochimica Polonica 45, 87-94 (1998)
  14. Wieczorek Z., Zdanowski K., Chlebicka L., Stępiński J., Jankowska M., Kierdaszuk B., Temeriusz A., Darżynkiewicz E. and Stolarski R., Fluorescence and NMR studies of intramolecular stacking of mRNA cap-analogues. Biochimica et Biophysica Acta 1354, 145-152 (1997)
  15. Kierdaszuk B., Gryczynski I. and Lakowicz J.R., Two-photon induced fluorescence of proteins. In Topics in Fluorescence Spectroscopy, Volume 5: Nonlinear and Two-Photon-Induced Fluorescence, (Lakowicz, J. Ed.) New York: Plenum Publishing Co., pp. 187-209 (1997)
  16. Kierdaszuk B., Modrak-Wojcik A. and Shugar D., Binding of phosphate and sulfate anions by purine nucleoside phosphorylase from E. coli: Ligand-dependent quenching of enzyme intrinsic fluorescence. Biophysical Chemistry 63, 107-118 (1997)
  17. Birnbaum K., Kierdaszuk B. and Shugar D., Crystal structure of 1,3,5-trimethyl-N4-hydroxycytosine, and its relevance to the mechanism of hydroxylamine mutagenesis. Nucleosides & Nucleotides 15, 1805-1819 (1996)
  18. Kierdaszuk B., Malak H. Gryczynski I., Calis P. and Lakowicz J.R., Fluorescence of reduced nicotinamides using one- and two-photon excitation. Biophysical Chemistry 62, 1-13 (1996)
  19. Lakowicz J.R., Kierdaszuk B., Malak H. and Gryczynski I., Fluorescence of horse liver alcohol dehydrogenase using one- and two-photon excitation. Journal of Fluorescence 6, 51-59 (1996)
  20. Lakowicz J.R., Kierdaszuk B., Calis P., Malak H. and Gryczynski I., Fluorescence anisotropy of tyrosine using one- and two-photon excitation. Biophysical Chemistry 56, 263-271 (1995)
  21. Krawiec K., Kierdaszuk B., Eriksson S., Munch-Petersen B. and Shugar D., Nucleoside triphosphate donors for nucleoside kinases: donor properties of UTP with human deoxycytidine kinase. Biochemical and Biophysical Research Communications 216, 42-48 (1995)
  22. Krawiec K., Kierdaszuk B. and Shugar D., 5'-Substituted 2'-Deoxycytidine as non-substrate Inhibitors of human deoxycytidine kinase. Nucleosides & Nucleotides 14, 495-499 (1995)
  23. Bretner M., Balinska M., Krawiec K., Kierdaszuk B., Shugar D. and Kulikowski T., Synthesis and Biological Activity of 5-Fluoro-2-thiocytosine Nucleosides. Nucleosides & Nucleotides 14, 657-660 (1995)
  24. Kierdaszuk B., Gryczynski I., Modrak-Wojcik A., Bzowska A., Shugar D. and Lakowicz J.R., Fluorescence of tyrosine and tryptophan in proteins using one- and two-photon excitation. Photochemistry and Photobiology 61, 319-324 (1995)

Lectures at international conferences, symposia and schools (1997-2003)

  1. "Emission Spectroscopy of Complex Formation between Enzymes and Identified Tautomeric and/or Ionic Species of Ligands." Fundamental Events in Photobiology, (Sariselka, Finland, 2002).
  2. "Enzyme-ligand interactions probed by induced tautomeric shift and enzyme-ligand FRET." 7th International Conference on methods and Applications of Fluorescence. Fluorescence Spectroscopy, Imaging and Probes. (Amsterdam, The Netherlands, 2001).
  3. "Interaction of Escherichia coli purine nucleoside phosphorylase (PNP) with formycin A (FA) probed by effect of induced tautomeric shifts and enzyme-ligand fluorescence resonance energy transfer on fluorescence and phosphorescence." 9th European Conference on the Spectroscopy of Biological Molecules (Prague, Czech Republic, 2001).
  4. "Multiphoton-induced fluorescence of biomolecules." International Conference on Photobiophysics in Technology and Medicine (Poznań, Poland, 2000).
  5. "Mechanism of action of purine nucleoside phosphorylases (PNP) by emission spectroscopy, X-ray crystallography and enzyme kinetics: Selective interaction of PNP with tautomeric and ionic species of substrates and inhibitors." International Conference on Enzymes of Nucleotide Metabolism - Structures, Catalysis and Drugs (Krogerup, Denmark, 2000).
  6. "Multiphoton-induced fluorescence of proteins and their constituents." An International Symposium "Protein 2000" - Celebrating a New Millennium of Protein Analysis, Synthesis and Function (London, England, 2000).
  7. "Enantiospecificity of human deoxycytidine kinases (dCK) and thymidine kinases (TK1, TK2) towards 3'-branched homologues of 2'-deoxynucleosides." 7th Symposium of European Society for the Study of Purine and Pyrimidine Metabolism in Man (Gdańsk, Poland, 1999).
  8. "Two-photon induced fluorescence of biomolecules." Laser Application in Life Sciences (Bratislava, Slovak Republic, 1998).
  9. "Mechanism of action of deoxycytidine kinase (dCK) and purine nucleoside phosphorylase (PNP) studied by fluorescence spectroscopy and enzyme kinetics." International Meeting on Enzyme Families in Nucleotide Metabolism (Krogerup Conference Center, Denmark, 1997).

List of selected posters, presented at international scientific meetings (1997-2002)

  1. Krawiec K., Kierdaszuk B., Kalinichenko E. N., Rubinova E. B., Mikhailopulo I. A., Eriksson S., Munch-Petersen B., Shugar D., Ability of adenosine-2'(3')-deoxy-3'(2')-triphosphates, and related analogues to replace ATP as phosphate donor for all four human, and the Drosophila melanogaster, deoxyribonucleoside kinases. XV International Round Table, Nucleosides, Nucleotides and Nucleic Acids, (Leuven, Belgium, 2002)
  2. Bochtler M., Szczepanowski R., Kierdaszuk B., Dandanell G., Shugar D., XapA, an atypical purine nucleoside phosphorylase. Deutsche Biophysical Society Meeting, (Germany, 2002)
  3. Rutkowska I, Sznee K., Nejman P., Stoychev G., Garstka M., Kierdaszuk B., Oxygen effect on the spectral properties of thylakoid membranes. 7th International Conference on Methods and Applications of Fluorescence. Fluorescence Spectroscopy, Imaging and Probes. (Amsterdam, The Netherlands, 2001)
  4. Włodarczyk J., Stoychev G., Kierdaszuk B., Specific interaction of E. coli purine nucleoside phosphorylase (PNP) with formycin A determined by its effects on fluorescence and phosphorescence anisotropy and enzyme-ligand fluorescence resonance energy transfer. 7th International Conference on Methods and Applications of Fluorescence. Fluorescence Spectroscopy, Imaging and Probes (Amsterdam, The Netherlands, 2001)
  5. Stoychev G., Kierdaszuk B., Shugar D., Interaction of E. coli purine nucleoside phosphorylase (PNP) with the cationic and zwitterionic forms of the fluorescent substrate N(7)-methylguanosine. 7th International Conference on Methods and Applications of Fluorescence. Fluorescence Spectroscopy, Imaging and Probes. (Amsterdam, The Netherlands, 2001)
  6. Włodarczyk J., Stoychev G., Kierdaszuk B., Interaction of Escherichia coli purine nucleoside phosphorylase (PNP) with formycin A (FA) probed by effect of induced tautomeric shift and enzyme <--> ligand fluorescence resonace energy transfer on fluorescence and phosphorescence. 7th International Conference on Methods and Applications of Fluorescence. Fluorescence Spectroscopy, Imaging and Probes. (Amsterdam, The Netherlands, 2001)
  7. Włodarczyk J., Wodzisławska M., Stoychev G., Koznakiewicz B., Shugar D., Kierdaszuk B., Formycin A and its N-methyl analogues, specific inhibitors of E. Coli purine nucleoside phosphorylase (PNP): Effect of induced tautomeric shifts and enzyme-ligand fluorescence and phosphorescence emission. International Conference on Photobiophysics in Technology and Medicine (Poznań, Poland, 2000)
  8. Modrak-Wójcik A., Stoychev G., Wierzchowski J., Shugar D. and Kierdaszuk B., Mechanism of action of purine nucleoside phosphorylases (PNP) by emission spectroscopy, X-ray crystallography and enzyme kinetics: Selective interaction of PNP with tautomeric and ionic species of substrates and inhibitors. International Conference on Enzymes of Nucleotide Metabolism - Structures, Catalysis and Drugs (Krogerup, Denmark, 2000)
  9. Modrak-Wójcik A., Kirilenko A., Shugar D., Kierdaszuk B., Effect of ionization of substrates and enzyme on phosphorolysis of N(7)-methylguanosine (m7Guo) by purine nucleoside phosphorylase (PNP). 7th International Symposium on Molecular Aspects of Chemotherapy, (Gdańsk, Poland, 1999)
  10. Zuberek J., Stojczew G., Munch-Petersen B., Shugar D., Kierdaszuk B., Transformation of dimeric low-affinity form of human thymidine kinase (TK1) to the tetrameric high-affinity form (ATP-form). 7th Symposium of European Society for the Study of Purine and Pyrimidine Metabolism in Man (Gdańsk, Poland, 1999)
  11. Kierdaszuk B., One and two-photon induced fluorescence of proteins. Methods and Applications of fluorescence spectroscopy (Paris, France, 1999)
  12. Kierdaszuk B., One and two-photon induced fluorescence of biomolecules. International Congress of Biophysics (New Delhi, India, 1999)
  13. Kierdaszuk B., Krawiec K., Kazimierczuk Z., Jacobsson U., Johansson N.G., Munch-Petersen B., Eriksson S. and Shugar D., Substrate/inhibitor specificities of human deoxycytidine kinase (dCK) and thymidine kinase (TK1 and TK2. 9th International/6th European Symposium on Purine and Pyrimidine Metabolism in Man (Gmunden, Austria, 1997).
  14. Krawiec K., Kierdaszuk B., Kalinichenko E.N., Mikhailopulo I.A., Shugar D., Unusual nucleoside triphosphate donors for nucleoside kinases: 3'-deoxyadenosine-2'-triphosphate and 2'-deoxyadenosine-3'-triphosphate. 6-th International Symposium on Molecular Aspects of Chemotherapy (Gdańsk, Poland, 1997)
  15. Nowak M., Lapinski L., Sobolewski A., Kiraga K., Kierdaszuk B., Photoinduced SYN-ANTI isomerization of N4-hydroxycytosines isolated in low-temperature matrices. XVIIIth International Conference on Photochemistry (Warsaw, Poland,1997)
  16. Kierdaszuk B., Gryczynski I., Malak H., Lakowicz J.R., Two-photon induced fluorescence of proteins. XVIIIth International Conference on Photochemistry (Warsaw, Poland, 1997)
  17. Modrak-Wójcik A., Shugar D., Kierdaszuk B., Termodynamic properties of purine nucleoside phosphorylase (PNP) from E. coli studied by temperature dependence of enzyme fluorescence and enzyme activity. XVIIIth International Conference on Photochemistry (Warsaw, Poland, 1997)
  18. Stojczew G., Shugar D., Kierdaszuk B., The pH-dependence of tyrosine fluorescence of E. coli purine nucleoside phosphorylase (PNP) and of enzyme-ligand (substrate) interactions. XVIIIth International Conference on Photochemistry (Warsaw, Poland, 1997)
  19. Kierdaszuk B., The scope and limitations for one- and two-photon induced fluorescence of protein systems. International Conference Femtochemistry III (Lund, Sweden, 1997)

Participation in research projects

Mechanism of action of purine nucleoside phosphorylases (PNP) based on spectroscopic, chromatographic, crystallographic and kinetic studies of enzyme-ligand complexes

Striking substrate/inhibitor specificities of four human, and the multifunctional Drosophila, 2'-deoxyribonucleoside kinases

Mechanism of function of thylakoid membranes in higher plants