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European Biophysics Journal

, Volume 14, Issue 1, pp 29–35 | Cite as

Fluorescence of 3-keto-steroids in aqueous solution

Probes for steroid-protein interactions
  • M. Kempfle
  • R. Müller
  • R. Palluk
  • K. A. Zachariasse
Article

Abstract

The physiologically important 3-keto-steroids are non-fluorescent or only weakly fluorescent in protic as well as in aprotic solvents. In contrast, the 4,6,8(14)-triene-3-one steroids are highly fluorescent in aqueous solution but they do not appreciably fluoresce in other solvents. Evidence is presented that the introduction of double bonds into the skeleton of the 3-keto-steroids leads to a decrease of the energy of the lowest π − π* state, bringing this level into the neighbourhood of the non-fluorescent n − π* state. As a consequence, for two states of approximately the same energy, relatively small perturbations such as those due to solvent interactions, protein binding and micelle formation, will then determine whether a system will fluoresce (π − π* state lowest) or not (n − π* state lowest). When the fluorescent 3-keto-steroids, having three conjugated double bonds, bind to proteins, the fluorescence intensity becomes almost zero, making these compounds useful as probes for steroid-protein interactions. This quenching of the fluorescence is explained by a decrease in energy of the n − π* state relative to the π − π* state of the steroids due to hydrophobic interactions with the proteins.

Key words

Fluorescent steroid probes steroid-protein interactions, energy alternation of n − π* and π − π* states (level crossing) 

Abbreviations

6,8-BDT

6,8-bisdehydrotestosterone; DMSO, dimethylsulfoxide

HPLC

high pressure liquid chromatography

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Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • M. Kempfle
    • 1
  • R. Müller
    • 1
  • R. Palluk
    • 1
  • K. A. Zachariasse
    • 2
  1. 1.Physiologisch-chemisches Institut der Universität BonnBonnFederal Republic of Germany
  2. 2.Abteilung SpektroskopieMax-Planck-Institut für biophysikalische ChemieGöttingenFederal Republic of Germany

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