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Quenching of the fluorescence of Tyr and Trp residues of firefly luciferase from Luciola mingrelica by the substrates

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Abstract

Luciferase of the firefly Luciola mingrelica is characterized by fluorescence of not only the unique Trp residue (λ em = 340 nm), but also that of Tyr residues (λ em = 308 nm). Quenching of the intrinsic fluorescence of the luciferase by its substrates luciferin and ATP (AMP) has been studied. Luciferin (LH2) quenches Trp fluorescence more efficiently than the fluorescence of Tyr residues. Two centers of quenching of Tyr fluorescence by ATP have been found corresponding apparently to the allosteric and active sites of the luciferase with K s(ATP) = 20 and 110 μM, respectively. The influence of one substrate on the affinity of luciferase to the second was investigated using fluorescence. ATP (AMP) binding to the allosteric sites of the luciferase significantly affects the affinity of luciferase to LH2. Formation of the complex between the luciferase and LH2 affects the affinity of both allosteric and active sites of the luciferase to ATP (AMP). The observed effects are probably connected with conformational changes in the luciferase molecule upon its interaction with the substrates.

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Abbreviations

K s :

dissociation constant of the luciferase complex with an effector

R 0 :

Forster radius (distance between donor and acceptor when efficiency of energy transfer is 50%)

K m :

Michaelis constant

LH2 :

luciferin

λ em, λ ex :

positions of the maxima of the fluorescence and excitation spectra, respectively

References

  1. Lakovich, J. (1986) Principles of Fluorescence Spectroscopy [Russian translation], Mir, Moscow.

    Google Scholar 

  2. Freifeld, D. (1980) Physical Biochemistry [Russian translation], Mir, Moscow.

    Google Scholar 

  3. McElroy, W. D., and De Luca, M. (1974) in Chemiluminescence and Bioluminescence (Cormier, M. J., Hercules, D. M., and Lee, J., eds.) Plenum Press, N. Y., pp. 285–311.

    Google Scholar 

  4. Sandalova, T. R., and Ugarova, N. N. (1999) Biochemistry (Moscow), 64, 962–967.

    CAS  Google Scholar 

  5. Dementieva, E. I., Brovko, L. Yu., Druzhinina, E. N., Gandelman, O. A., and Ugarova, N. N. (1986) Biokhimiya, 51, 130–139.

    Google Scholar 

  6. Gandelman, O. A., Brovko, L. Yu., Ugarova, N. N., and Shchegolev, A. A. (1990) Biokhimiya, 55, 1052–1058.

    CAS  Google Scholar 

  7. Chudinova, E. A., Dementieva, E. I., Brovko, L. Yu., Savitskii, A. P., and Ugarova, N. N. (1999) Biochemistry (Moscow), 64, 1097–1103.

    CAS  Google Scholar 

  8. Brovko, L. Yu., Demenieva, E. I., Gandelman, O. A., Chikishev, A. Yu., Shkurinov, A. P., and Ugarova, N. N. (1997) J. Fluoresc., 7, 139–142.

    Google Scholar 

  9. Cherednikova, E. Yu., Chikishev, A. Yu., Dementieva, E. I., Kossobokova, O. V., and Ugarova, N. N. (2001) J. Photochem. Photobiol. B: Biol., 60, 7–11.

    Article  CAS  Google Scholar 

  10. Dementieva, E. I., Fedorchuk, E. A., Brovko, L. Yu., Savitskii, A. P., and Ugarova, N. N. (2000) Biosci. Rep., 20, 21–30.

    Article  PubMed  CAS  Google Scholar 

  11. Nojioma, H., Ikai, A., and Noda, H. (1976) Biochim. Biophys. Acta, 27, 20–27.

    Google Scholar 

  12. Schlessinger, J., Roche, R. S., and Steinberg, I. Z. (1975) Biochemistry, 14, 255–261.

    Article  PubMed  CAS  Google Scholar 

  13. Sackett, D. L., Ruvinov, S. B., and Thompson, J. (1999) Protein Sci., 8, 2121–2129.

    Article  PubMed  CAS  Google Scholar 

  14. Ruan, K., Li, J., Liang, R., Xu, C., Yu, Y., Lange, R., and Balny, C. (2002) Biochem. Biophys. Res. Commun., 293, 593–597.

    Article  PubMed  CAS  Google Scholar 

  15. Yang, Y., Shao, Z., Chen, X., and Zhou, P. (2004) Biomacromolecules, 5, 773–779.

    Article  PubMed  CAS  Google Scholar 

  16. Finazzi-Agro, A., Rotilio, G., Avigliano, L., Guerrieri, P., Boffi, V., and Mondovi, B. (1970) Biochemistry, 9, 2009–2014.

    Article  PubMed  CAS  Google Scholar 

  17. Pongs, O. (1970) Biochem. Biophys. Res. Commun., 38, 431–437.

    Article  PubMed  CAS  Google Scholar 

  18. Conti, E., Francs, N. P., and Brick, P. (1996) Structure, 4, 287–289.

    Article  PubMed  CAS  Google Scholar 

  19. Ugarova, N. N. (1989) J. Biolum. Chemilum., 4, 406–418.

    Article  CAS  Google Scholar 

  20. Talebarovskaya, I. K., Katkova, V. A., Ryzhova, V. V., and Shchegolev, A. A. (1983) RF Certificate of Authorship No. 1192324.

  21. Lundovskich, L. A., Leontieva, O. V., Dementieva, E. I., and Ugarova, N. N. (1999) in Bioluminescence and Chemiluminescence. Perspectives for 21st Century (Stanley, P. E., ed.) John Wiley, Chichester, pp. 420–424.

    Google Scholar 

  22. Filipova, N. Yu., Dukhovich, A. F., Bukatina, V. A., and Ugarova, N. N. (1984) Biokhimiya, 49, 1965–1971.

    Google Scholar 

  23. DeLuca, M., and McElroy, W. D. (1984) Biochem. Biophys. Res. Commun., 123, 764–770.

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Faculty of Chemistry, Lomonosov Moscow State University, 119992, Moscow, Russia

    T. N. Vlasova & N. N. Ugarova

Authors
  1. T. N. Vlasova
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  2. N. N. Ugarova
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Correspondence to T. N. Vlasova.

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Original Russian Text © T. N. Vlasova, N. N. Ugarova, 2007, published in Biokhimiya, 2007, Vol. 72, No. 9, pp. 1182–1188.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM07-108, August 19, 2007.

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Vlasova, T.N., Ugarova, N.N. Quenching of the fluorescence of Tyr and Trp residues of firefly luciferase from Luciola mingrelica by the substrates. Biochemistry Moscow 72, 962–967 (2007). https://doi.org/10.1134/S0006297907090064

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  • DOI: https://doi.org/10.1134/S0006297907090064

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