DOI QR코드

DOI QR Code

Critical Role of Glu175 on Stability and Folding of Bacterial Luciferase: Stopped-flow Fluorescence Study

  • Shirazy, Najmeh Hadizadeh (Department of Biophysics & Biochemistry, Faculty of sciences, Tarbiat Modares University) ;
  • Ranjbar, Bijan (Department of Biophysics & Biochemistry, Faculty of sciences, Tarbiat Modares University) ;
  • Hosseinkhani, Saman (Department of Biophysics & Biochemistry, Faculty of sciences, Tarbiat Modares University) ;
  • Khalifeh, Khosrow (Department of Biophysics & Biochemistry, Faculty of sciences, Tarbiat Modares University) ;
  • Madvar, Ali Riahi (Department of Biophysics & Biochemistry, Faculty of sciences, Tarbiat Modares University) ;
  • Naderi-Manesh, Hossein (Department of Biophysics & Biochemistry, Faculty of sciences, Tarbiat Modares University)
  • Published : 2007.07.31

Abstract

Bacterial luciferase is a heterodimeric enzyme, which catalyzes the light emission reaction, utilizing reduced FMN (FMNH2), a long chain aliphatic aldehyde and $O_2$, to produce green-blue light. This enzyme can be readily classed as slow or fast decay based on their rate of luminescence decay in a single turnover. Mutation of Glu175 in $\alpha$ subunit to Gly converted slow decay Xenorhabdus Luminescence luciferase to fast decay one. The following studies revealed that changing the luciferase flexibility and lake of Glu-flavin interactions are responsible for the unusual kinetic properties of mutant enzyme. Optical and thermodynamics studies have caused a decrease in free energy and anisotropy of mutant enzyme. Moreover, the role of Glu175 in transition state of folding pathway by use of stopped-flow fluorescence technique has been studied which suggesting that Glu175 is not involved in transition state of folding and appears as surface residue of the nucleus or as a member of one of a few alternative folding nuclei. These results suggest that mutation of Glu175 to Gly extended the structure of Xenorhabdus Luminescence luciferase, locally.

Keywords

Anisotropy;Bacterial luciferase;Stopped-flow fluorescence;$\varphi$-value

References

  1. Anil, B., Sato, S., Cho, J.-H. and Raleigh, D. P. (2005) Fine structure analysis of a protein folding transition state; distinguishing between hydrophobic stabilization and specific packing. J. Mol. Biol. 354, 693-705. https://doi.org/10.1016/j.jmb.2005.08.054
  2. Bradford, M. M. (1976) A rapid and sensitive method for the quantization of microgram quantities of protein utilizinf the principle of protein -dye binding. Anal. Biochem. 72, 248-254. https://doi.org/10.1016/0003-2697(76)90527-3
  3. Cline, T. W. and Hasting, J. W. (1974) Mutated luciferase with altered bioluminescence emission spectra. J. Biol. Chem. 294, 4668-4669.
  4. Evans, M. G. and Polanyi, M. (1935) Some applications of the transition state method to the calculation of reaction velocities, especially in solution. Trans. Faraday Soc. 31, 875-894. https://doi.org/10.1039/tf9353100875
  5. Eyring, H. (1935) The activated complex and the absolute rate of chemical reactions. Chem. Rev. 17, 65-77. https://doi.org/10.1021/cr60056a006
  6. Fasman, G. D. (1996) Circular Dichroism and conformational analysis of biomolecules Plenum. Press. N. Y.
  7. Fersht, A. R., Matoaschek, A. and Serrano, L. (1992) The folding of an enzyme. I. theory of protein engineering analysis of stability and pathway of protein folding. J. Biol. 224, 771-782. https://doi.org/10.1016/0022-2836(92)90561-W
  8. Finkelstein, A. V. and Petitsyn, O. B. (2002) Protein physics. A course of lectures, part V: Cooperative transition in protein inolecules.pp. 260-261. Academic press, London, UK.
  9. Fisher, A. J., Thompson, T. B., Thoden, J. B., Baldwin, T. O. and Reyment I. (1996) The $1.5{\AA}$ resolution crystal structure of bacterial luciferase in low salt condition. J. Bio. Chemistry 271, 21956-21968. https://doi.org/10.1074/jbc.271.36.21956
  10. Gibson, Q. H. and Hasting, J. W. (1962) The oxidation of reduced flavin mononucleotide by molecular oxygen. Biochem. J. 83, 368-377. https://doi.org/10.1042/bj0830368
  11. Gunsalus-Miguel, A. Meighen, E. A., Nicoli, M. Z., Nealson, K. H. and Hastings, J. W. (1972) Purification and properties of bacterial luciferase. J. Biol. Chemistry 247, 398-404.
  12. Hastings, J. W. and Gibson, Q. H. (1963) Intermediate in the bioluminescence oxidation of reduced flavin mononucleotide. J. Biol. Chemistry 238, 2537-2554.
  13. Hosseinkhani, S., Ranjbar, B., Naderi-Manesh, H. and Nemat-Goegani, M. (2004) Chemical modification of glucose oxidase:possible formation of molten globule-like intermediate structure. FEBS Lett. 561, 213-216. https://doi.org/10.1016/S0014-5793(04)00134-6
  14. Hosseinkhani, S., Szittner, R. and Meighen, E. A. (2005) Random mutagenesis of bacterial luciferase: Critical role of $Glu^{175}$ in the control of luminescence decay. Biochem. J. 385, 1-6. https://doi.org/10.1042/BJ20041107
  15. Kocks, U. F., Tome, C. N. and Wenk, H.-R. (1998) Texture and anisotropy. Chapter 1: Anisotropy and symmetry. pp. 11-12. Cambridge University Press, Cambridge, UK.
  16. Lin, L. Y., Sulea, T., Szittner, R., Vassilyev, V., Purisima, E. O. and Meighen, A. (2001) Modeling of bacterial luciferase-flavin mononucleotide complex combining flexible docking with structure-activity data. Protein Sci. 10, 1563-1571. https://doi.org/10.1110/ps.7201
  17. Meighen, E. A. and Mackenzie, R. E. (1973) Flavin specifity of enzyme-substrate intermediates in the bacterial bioluminescence reaction. Biochemistry 12, 1482-1491. https://doi.org/10.1021/bi00732a003
  18. Nealson, K. H. (1978) Isolation, identification and manipulation of luminous bacteria. Methods Enzymol 57, 153-166. https://doi.org/10.1016/0076-6879(78)57017-1
  19. Nealson, K. H. and Hasting, J. W. (1979) Isolation, identification and manipulation of luminous bacteria. Microbial. Rev. 43, 496-518.
  20. Privalov, P. L. (1979) Stability of proteins. Adv. Protein Chemistry 33, 167-236. https://doi.org/10.1016/S0065-3233(08)60460-X
  21. Protasevich, I., Ranjbar, B., Lobachov, V., Makarov, A., Gilli, R., Briand, C., Lafitte, D. and Haiech, J. (1997) Conformation and thermal denaturation of apocalmodulin: role of electrostatic mutations. Biocheistry 36, 2017-2024. https://doi.org/10.1021/bi962538g
  22. Riahi Madvar, A., Hosseinkhani, S., Khajeh, Kh., Ranjbar, B. and Asoodeh, A. (2005) Implication of a critical residue (Glu 175) in structure and function of bacterial luciferase. FEBS Lett. 579, 4701-4706. https://doi.org/10.1016/j.febslet.2005.07.040
  23. Schippers, P. H. and Dekkers, H. (1981) Direct determination of absolute circular Dichroism data and calibration of commertioal instrument. Anal. Chem. 53, 778-788. https://doi.org/10.1021/ac00229a008
  24. Serrano, L., James, T., Kellis, Jr., Cann, P., Matouschek, A. and Fersht, A. R. (1992). The folding of an enzyme: II. Substructure of barnase and contribution of different interactions to protein stability. J. Mol. Biol. 224, 783-804. https://doi.org/10.1016/0022-2836(92)90562-X
  25. Serrano, L., Kellis, J. T., Cann.P., Matouschek, A. and Fersht, A. (1992) The folding of an enzyme.II.Substract of barnase and the contribution of different interactions to protein stability. J. Mol. Biol. 224, 783-804. https://doi.org/10.1016/0022-2836(92)90562-X
  26. Serrano, L., Matouschek, A. and Fersht, A. R. (1992) The folding of enzyme.III.Structure of transition state for unfolding of barnase analyzed by protein engineering procedure. J. Mol. Biol. 224, 805-818. https://doi.org/10.1016/0022-2836(92)90563-Y
  27. Szittner, R. and Meighen, E. (1990) Nucleotide sequence, expression and properties of luciferase coded by lux genes from a terrestrial bacterium. J. Biol. Chemistry 265, 16581-16587.
  28. Szittner, R. and Meighen, E. A. (1993) Subunit interactions and the role of the lux polypeptide in controlling thermal stability and catalytic properties in recombinant luciferase hybrids. Biochem. Biophys. Acta 1158, 137-145. https://doi.org/10.1016/0304-4165(93)90007-U
  29. Takakuwa, T., Konno, T. and Meguro, H. A. (1985) New standard substance for calibration of circular dichroism: ammonium d-10-camphorsulfonate. Anal. Sci. 1, 215-225. https://doi.org/10.2116/analsci.1.215
  30. Urray, D. W. (1985) Absorption, circular Dichroism and optical rotatory dispersion of polypeptides,proteins,prosthetic group and biomembrane. Mod. Phys. biochemistry part A, 175-345.

Cited by

  1. Hydrophobin-1 promotes thermostability of firefly luciferase vol.283, pp.13, 2016, https://doi.org/10.1111/febs.13757
  2. Roles of trehalose and magnesium sulfate on structural and functional stability of firefly luciferase vol.62, pp.2, 2010, https://doi.org/10.1016/j.molcatb.2009.09.015
  3. Relationship between stability and bioluminescence color of firefly luciferase vol.9, pp.3, 2010, https://doi.org/10.1039/b9pp00161a
  4. Spatio-Temporal Control of Bacterial-Suspension Luminescence Using a PDMS Cell vol.43, pp.11, 2010, https://doi.org/10.1252/jcej.10we137
  5. Bioluminescence intensity difference observed in luminous bacteria groups with different motility vol.48, pp.3, 2009, https://doi.org/10.1111/j.1472-765X.2008.02525.x
  6. Circular Dichroism Techniques: Biomolecular and Nanostructural Analyses- A Review vol.74, pp.2, 2009, https://doi.org/10.1111/j.1747-0285.2009.00847.x
  7. Structural distinctions of fast and slow bacterial luciferases revealed by phylogenetic analysis vol.32, pp.20, 2016, https://doi.org/10.1093/bioinformatics/btw386