Skip to main content

Advertisement

Log in

Characterization of human immunodeficiency virus-1 (HIV-1) rev by (time-resolved) fluorescence spectroscopy

  • Fluorescence Imaging and Microscopy
  • Published:
Journal of Fluorescence Aims and scope Submit manuscript

Abstract

Fluorescence spectroscopy has been applied to the single tryptophan-containing regulatory protein Rev of human immunodeficiency virus (HIV-1). The fluorescence emission was found to have a maximum at 336 nm which refers to a surrounding of the chromophore of intermediate polarity. Fluorescence transients recorded at the maximum of fluorescence were found to decay nonexponentially. A bimodal lifetime distribution is obtained from exponential series analysis (ESM) with centers at 1.7 and 4.5 ns. Two microenvironments for tryptophan are suggested to be responsible for the two lifetime distributions. No innerfilter effect occurred in a Rev solution up to a concentration of 40 μM. A data quality study of ESM analysis as function of collected counts in the peak channel maximum (CIM) showed that, for reliable reconvolution, at least 15,000 CIM are necessary. The widths of the two distributions are shown to be temperature dependent. The broadening of the lifetime distributions when the temperature is raised to 50°C is interpreted as extension of the number of conformational substates which do not interconvert on the fluorescence time scale. The thermal deactivation (temperature quenching) is reflected in a constant decrease in the center of the short-lived lifetime distribution.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. J. Sodroski, W. C. Goh, C. Rosen, A. Dayton, E. Terwilliger, and W. Haseltine (1986)Nature 321, 412–417.

    PubMed  Google Scholar 

  2. S. K. Arya, C. Guo, S. F. Josephs, and F. Wong-Staal (1985)Science 229, 69–73.

    PubMed  Google Scholar 

  3. E. T. Dayton, D. M. Powell, and A. I. Dayton (1989)Science 246, 1625–1629.

    PubMed  Google Scholar 

  4. E. Terwilliger, R. Burghoff, R. Sia, J. Sodroski, W. Haseltine, and C. Rosen (1988).J Virol 62, 655–658.

    PubMed  Google Scholar 

  5. T. J. Daly, K. S. Cook, G. S. Gray, T. E. Maione, and J. R. Rusche (1989)Nature 342, 816–819.

    PubMed  Google Scholar 

  6. M. L. Zapp and M. R. Green (1989)Nature 342, 714–716.

    PubMed  Google Scholar 

  7. A. W. Cochrane, C.-H. Chen, and C. A. Rosen (1990)Proc. Natl. Acad. Sci. 87, 1198–1202.

    PubMed  Google Scholar 

  8. S. Heaphy J. T., Finch, M. J. Gait, J. Karn, and M. Singh (1991)Proc. Natl. Acad. Sci. USA 88, 7366–7370.

    PubMed  Google Scholar 

  9. M. H. Malim, L. S. Tiley, D. F. McCarn, J. R. Rusche, J. Hauber, and B. R. Cullen (1990)Cell 60, 675–683.

    PubMed  Google Scholar 

  10. H. S. Olsen, P. Nelböck, A. W. Cochrane, and C. A. Rosen (1990)Science 247, 845–848.

    PubMed  Google Scholar 

  11. M. H. Malim, S. Böhnlein, J. Hauber, and B. R. Cullen (1989)Cell 58, 205–214.

    PubMed  Google Scholar 

  12. B. Mermer, B. K. Felber, M. Campbell, and G. N. Pavlakis (1990)Nucleic Acids Res. 18, 2037–2044.

    PubMed  Google Scholar 

  13. L. K. Venkatesh and G. Chinnadurai (1990)Virology 178, 327–330.

    PubMed  Google Scholar 

  14. T. J. Hope, X. Huang, D. McDonald, and T. G. Parslow (1990).Proc. Natl. Acad. Sci. USA 87, 7787–7791.

    PubMed  Google Scholar 

  15. M. H. Malim, D. F. McCarn, L. S. Tiley, and B. R. Cullen (1991)J. Virol. 65, 4248–4254.

    PubMed  Google Scholar 

  16. I. Weichselbraun, G. K. Farrington, J. R. Rusche, E. Böhnlein, and J. Hauber (1992)J. Virol. 66, 2583–2587.

    PubMed  Google Scholar 

  17. D. Bevec, M. Dobrovnik, J. Hauber, and E. Boehnlein (1992).Proc. Natl. Acad. Sci. USA 89, 9870–9874.

    PubMed  Google Scholar 

  18. M. H. Malim, W. W. Freimuth, J. Liu, T. J. Boyle, K. Lyerly, B. R. Cullen, and G. J. Nabel (1992)J. Exp. Med. 176, 1197–1201.

    PubMed  Google Scholar 

  19. M. Auer, H.-U. Gremlich, J. M. Seifert, T. J. Daly, T. G. Parslow, G. Casari, and H. Gstach (1993)Biochemistry 33, 2988–2996.

    Google Scholar 

  20. T. P. Wingfield, S. J. Stahl, M. A. Payton, S. Venkatesan, M. Misra, and A. A. C. Steven (1991)Biochemistry 31, 7527.

    Google Scholar 

  21. M. C. Nalin, R. D. Purcell, D. Antelman, D. Mueller, L. Tomchak, B. Wegrzynski, E. McCorney, V. Toome, R. Kramer, and M.-C. Hsu (1990)Proc. Natl. Acad. Sci. USA 87, 7593–7597.

    PubMed  Google Scholar 

  22. M. R. Eftink (1991)Methods Biochem. Anal. 35, 127–205.

    PubMed  Google Scholar 

  23. G. Landl, T. Langthaler, H. W. Engl, and H. F. Kauffmann (1991)J. Comp. Phys. 95, 1–28.

    Google Scholar 

  24. T. J. Daly, R. D. Doten, P. Rennert, M. Auer, H. Jaksche, A. Donner, G. Fisk, and J. R. Rusche (1993)Biochemistry 32, 10497–10505.

    PubMed  Google Scholar 

  25. T. J. Daly, P. Rennert, P. Lynch, J. Barry, M. Dundas, J. R. Rusche, R. C. Doten, M. Auer, and G. K. Farrington (1993)Biochemistry 32, 8945–8954.

    PubMed  Google Scholar 

  26. E. Prenner, A. Hermetter, G. Landl, H. Stuetz, H. F. Kauffman, and A. J. Kungl (1993)J. Phys. Chem. 97, 2788–2792.

    Google Scholar 

  27. M. K. Hong, D. Braunstein, B. R. Crown, H. Frauenfelder, I. E. Iben, J. R. Mourant, P. Ormos, R. Scholl, A. Schulte, P. J. Steinbach, A. Xie, and R. D. Young (1990)Biophys. J. 58, 429–436.

    PubMed  Google Scholar 

  28. E. Gratton, J. R. Alcala, and F. G. Prendergast (1989) in D. M. Jameson, G. D. Reinhart (Eds.),Fluorescent Biomolecules, Plenum Press, New York/London, pp. 17–32.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kungl, A.J., Seidel, C., Schilk, A. et al. Characterization of human immunodeficiency virus-1 (HIV-1) rev by (time-resolved) fluorescence spectroscopy. J Fluoresc 4, 299–302 (1994). https://doi.org/10.1007/BF01881444

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01881444

Key Words

Navigation