Skip to main content
Log in

Detecting enzymes in living cells using fluorogenic substrates

  • Probe Development
  • Published:
Journal of Fluorescence Aims and scope Submit manuscript

Abstract

Characteristics of fluorogenic substrates designed for detection of enzyme activity in living cells are reviewed. Improved retention of the fluorescent products in the cell of origin can be achieved by structural modifications to the substrate that result in association with membrane lipids or conjugation to intracellular glutathione. Newly-developed substrates that yield fluorescent precipitates provide the additional advantage of allowing subcellular localization of sites of enzymatic activity. Improved detection sensitivity can also be achieved by targeted delivery of substrates for processing by specific organelles. Substrates designed for monitoring oxidative activity and lipid metabolism provide examples of this approach.

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

Access this article

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. G. G. Guilbault (1990) in G. G. Guilbault (Ed.),Practical Fluorescence, 2nd ed., Marcel Dekker, New York, pp. 683–673.

    Google Scholar 

  2. M. Manafi, W. Kneifel, and S. Bascomb (1991)Microbiol. Rev.,55, 335–348.

    PubMed  Google Scholar 

  3. G. P. Nolan, S. N. Fiering, J.-F. Nicolas, and L. A. Herzenberg (1988)Proc. Natl. Acad Sci. U.S.A. 85, 2603–2607.

    PubMed  Google Scholar 

  4. S. N. Fiering, M. Roederer, G. P. Nolan, D. R. Micklem, D. R. Parks, and L. A. Herzenberg (1991)Cytometry 12, 291–301.

    PubMed  Google Scholar 

  5. Y.-Z. Zhang, J. J. Naleway, K. D. Larison, Z. Huang, and R. P. Haugland (1991)FASEB J. 5, 3108–3113.

    PubMed  Google Scholar 

  6. M. B. Gu, P. Todd, and D. S. Kompala (1993)Biotechnol. Bioeng. 42, 1113–1123.

    Google Scholar 

  7. B. G. Rosser, S. P. Powers, and G. J. Gores (1993)J. Biol. Chem. 268, 23593–23600.

    PubMed  Google Scholar 

  8. F. Dolbeare and M. Vanderlaan (1979).J. Histochem. Cytochem. 27, 1493–1495.

    PubMed  Google Scholar 

  9. R. B. Banati, G. Rothe, G. Valet, and G. W. Kreutzberg (1993)Glia 7, 183–191.

    PubMed  Google Scholar 

  10. G. Rothe, I. Assfalg-Machleidt, W. Machleidt, S. Klingel, C. Zirkelbach, R. B. Banati, W. F. Mangel, and G. Valet (1992).Biol. Chem. Hoppe-Seyler 373, 547–554.

    PubMed  Google Scholar 

  11. W. H. Fishman, Y. Nakajima, C. Anstiss, and S. Green (1964)J. Histochem. Cytochem. 12, 298–305.

    PubMed  Google Scholar 

  12. E. J. M. Speel, B. Schutte, J. Wiegant, F.C.S. Ramaekers, and A. H. N. Hopman (1992)J. Histochem. Cytochem. 40, 1299–1308.

    PubMed  Google Scholar 

  13. T. C. Ryan, G. J. Weil, P. E. Newburger, R. P. Haugland, and E. R. Simons (1990)J. Immunol. Methods 130, 223–233.

    PubMed  Google Scholar 

  14. G. Rothe, A. Oser, and G. Valet (1988)Naturwissenschaften 75, 354–355.

    PubMed  Google Scholar 

  15. G. Rothe, A. Emmendörffer, A. Oser, J. Roesler, and G. Valet (1991)J. Immunol. Methods 138, 133–135.

    PubMed  Google Scholar 

  16. G. Rothe and G. Valet (1990)J. Leukocyte Biol. 47, 440–448.

    PubMed  Google Scholar 

  17. R. A. Prough, M. D. Burke, and R. T. Mayer (1978)Methods Enzymol. 52, 372–377.

    PubMed  Google Scholar 

  18. R. E. Pagano, O. C. Martin, H. C. Kang, and R. P. Haugland (1991)J. Cell Biol.,113, 1267–1279.

    PubMed  Google Scholar 

  19. J. Kasurinen (1992)Biochem. Biophys. Res. Comm. 187, 1594–1601.

    PubMed  Google Scholar 

  20. T. Meshulam, H. Herscovitz, D. Casavant, J. Bernado, R. Roman, R. P. Haugland, G. S. Strohmeier, R. D. Diamond, and E. R. Simons (1992)J. Biol. Chem. 267, 21465–21470.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Haugland, R.P., Johnson, I.D. Detecting enzymes in living cells using fluorogenic substrates. J Fluoresc 3, 119–127 (1993). https://doi.org/10.1007/BF00862728

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00862728

Key Words

Navigation