BMB Faculty

John Lee

Professor of Biochemistry and Molecular Biology
Ph.D. (1960) University of New South Wales
Post-doctoral (1961-63) Johns Hopkins University

President (2002-2004) International Society for Bioluminescence and Chemiluminescence: www.unibo.it/isbc/

   STRUCTURE AND MECHANISM OF BIOLUMINESCENT  PROTEINS     
     

     The major research interest is in the biophysical aspects of bioluminescence.  We utilize picosecond fluorescence dynamics and macromolecular structural methods: high-field nuclear magnetic resonance and X-ray crystallography.  The properties of certain fluorescent proteins occurring in the bioluminescent bacteria have been given detailed study, such as spectral properties, primary sequence, and three-dimensional structure. Current work concentrates on the calcium-regulated photoproteins, obelin and aequorin.  These studies involve collaborations with groups in the U.S., Holland and Russia.
 

 Full publications : 149

Recent representative publications:

  •   Petushkov, V., Gibson, B. G., and Lee, J.  Direct measurement of excitation transfer in the protein complex of bacterial luciferase hydroxyflavin and the associated yellow fluorescence proteins from Vibrio fischeri Y-1. Biochemistry  35:8413-8418 (1996). 
  •   Lee, J. Bioluminescence.  In, "Encyclopedia of Life Sciences", www.els.com (Macmillan, London) (1999). 
  •   Vysotski ES, Liu ZJ, Rose J, Wang BC, Lee J.. Preparation and preliminary study of crystals of the calcium-regulated photoprotein obelin from the bioluminescent hydroid  Obelia longissima.  Acta Crystallogr D55: 1965-1966 (1999). 
  • Vysotski ES, Liu ZJ, Rose J, Wang BC, Lee J.  Preparation and X-ray crystallographic analysis of recombinant obelin crystals diffracting to beyond 1.1 Å.  Acta Crystallogr D57: 1919 (2001). 
  • Liu ZJ, Vysotski ES, Chen CJ, Rose J, Lee J, Wang BC. (2000). Structure of the Ca2+-regulated photoprotein obelin at 1.7 Å resolution determined directly from its sulfur substructure. Protein Sci 9: 2085 (2000). 
  • Deng L, Vysotski ES, Liu Z-J, Markova S, Malikova NP, Lee J, Rose J, Wang B-C. Structural basis for the emission of violet bioluminescence from a W92F obelin mutant. FEBS Lett. 506: 281(2001).
  • Markova SV, Vysotski ES, Blinks JR, Burakova LP, Wang B-C, Lee J. Obelin from the marine hydroid Obelia geniculata: cloning, expression, and comparison of some properties with those of other Ca2+-regulated photoproteins. Biochemistry 41: 2227 (2002)..
  • Svetlana V. Markova, Eugene S.Vysotski, John R. Blinks,Ludmila P. Burakova, B-C. Wang, and John Lee. (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.
  • Eugene S. Vysotski, Zhi-Jie Liu, Svetlana V. Markova, John R. Blinks, Lu Deng, Ludmila A. Frank, Michelle Herko, Natalia P. Malikova, John P. Rose, Bi-Cheng Wang, and John Lee. (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.
  • Lee, J., and Vysotski, E.S. (2003). Structure and spectra in bioluminescence. In, "The Digital Photobiology Compendium" (Valenzeno, D., Ed.)http://www.photobiology.info.
  • VN Petushkov, IHM van Stokkum, B. Goberts, F. van Mourik, J. Lee, R van Grondelle, and AJWG Visser (2003). Ultrafast fluorescence relaxation spectroscopy of lumazine and riboflavin free and bound to antenna proteins from bioluminescent bacteria.  J. Phys. Chem. 107: 10934-10939.
  • NP Malikova, GA Stepanyuk, LA Frank, SV Markova, ES Vysotski, and J. Lee.  (2003) Spectral tuning of obelin bioluminescence by mutations of Trp92. FEBS Lett. 554:184-188.
  • Z-J Liu, ES Vysotski, L Deng, J Lee, J Rose, and BC Wang. (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.
  • L Deng, SV Markova, ES Vysotski, Z-J Liu, J Lee, J Rose, and B-C Wang. (2004) Preparation and X-ray crystallographic analysis of the Ca2+-discharged photoprotein obelin. Acta Cryst. D60: 512-514.
  • ES Vysotski and J Lee. (2004) Ca2+-regulated photoproteins: Structural insight into the bioluminescence mechanism.  Acc. Chem. Res. 37: 405-415.
  • L Deng, SV Markova , ES Vysotski,  Z-J Liu, J Lee, J Rose and B-C Wang (2004). Crystal structure of a Ca2+-discharged photoprotein: Implications for the mechanisms of the calcium trigger and the bioluminescence. J. Biol. Chem. 279: 33647-52.
  • L Deng, SV Markova , ES. Vysotski,  Z-J Liu, J Lee, J Rose and B-C Wang (2005). All three Ca2+-binding loops of photoproteins bind calcium ion: The crystal structures of calcium-loaded apo-aequorin and apo-obelin. Protein Science 14: 663-675.
  •  GA Stepanyuk, S Golz, SV Markova, LA Frank, J Lee, ES Vysotski.(2005). Interchange of aequorin and obelin bioluminescence color is determined by substitution of one active site residue of each photoprotein. FEBS Lett. 579: 1008-1114.
  •  Zhi-Jie Liu, Galina A. Stepanyuk, Eugene S. Vysotski, John Lee, Svetlana V. Markova, Natalia P. Malikova, and Bi-Cheng Wang. (2006). Crystal structure of obelin following Ca2+ triggered bioluminescence suggests neutral coelenteramide as the primary excited state. Proc. Natl. Acad. Sci. USA 103:2570-2575.