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.
Similar content being viewed by others
Abbreviations
- 6,8-BDT:
-
6,8-bisdehydrotestosterone; DMSO, dimethylsulfoxide
- HPLC:
-
high pressure liquid chromatography
References
Becker RS, Kasha M (1955) Luminescence spectroscopy of porphyrin-like molecules including the chlorophylls. J Am Chem Soc 77:3669–3670
Birks JB (1977) In: Mielenz KD, Velapoldi RA, Mavrodineanu R (eds) Standardisation in spectrometry and luminescence measurements National Bureau of Standards, Washington
Bredereck K, Förster Th, Oesterlin HG (1962) In: Kallmann HP, Spruch GM (eds) Luminescence of organic and inorganic materials. Wiley, New York, pp 161–175
Chen M, Grätzel M, Thomas JK (1975) Kinetic studies in bile acid micelles. J Am Chem Soc 97:2052–2057
Dederen JC, Coosemans L, De Schryver FC, Van Dormael A (1979) Complex solvent dependence of pyrenealdehyde fluorescence as a micellar polarity probe. Photochem Photobiol 30:443–447
Demas JN, Crosby GA (1971) The measurement of photoluminescence quantum yields. A review. J Phys Chem 75: 991–1024
Inuzuka S, Becker RS (1974) Theoretical consideration of lower excited states of trans-polyene carbaldehydes. I. Electronic structure and oscillator strength of π − π * transition. Bull Chem Jpn 47:88–91
Inuzuka S (1975) Theoretical consideration of lower excited states of trans-polyene carbaldehydes. II. Radiative and non-radiative properties. Bull Chem Soc Jpn 48:779–782
Kasha M (1949) Characterization of electronic transitions in complex molecules. Disc Faraday Soc 9:14–19
Kempfle M, Müller R, Palluk R (1984) Fluoreszierende Steroide-Wechselwirkungen mit Cyclodextrinen Fresenius Z Anal Chem 317:700–701
Kempfle M, Breuer H, Müller R, Palluk R (1985) Fluorescent steroids and their interactions with 3α-and β-hydroxysteroid dehydrogenases. Acta Endocrinol Suppl 108: 112–113
Mataga N, Kubota K (1970) Molecular interactions and electronic spectra. Marcel Dekker, New York, p 355
Müller R, Palluk R, Kempfle M (1982) Charakterisierung der Fluoreszenzeigenschaften von 4,6,8(14)-Trien-3-on-Steroiden. Fresenius Z Anal Chem 311:369–370
Müller R, Palluk R, Kempfle M (1983) 4,6,8(14)-trien-3-one steroids as fluorescing components in preparations of 4,6-diene-3-one steroids. Arch Pharm, 316:91–92
Müller R, Palluk R, Ehlenz K, Kempfle M (1985) Fluorescent steroids: Homogeneous fluorescence immunoassay of cortisol in plasma by means of 17α-hydroxy-4,6,8(14) pregnatriene-3,20-dione. Acta Endocrinol Suppl 108:115–116
Müller R, Palluk R, Kempfle M (1986) Fluoreszierende 4,6,8(14)-Trien-3-on Steroide als Indikatoren zur Untersuchung der Proteinbindung von Steroiden. Stud Biophys 109 (in press)
Palluk R (1983) Protein-Wechselwirkungen fluoreszierender Steroide. Thesis, University of Bonn
Palluk R, Müller R, Kempfle M (1982) Untersuchung von Steroid-Protein-Wechselwirkungen mit Hilfe der Fluoreszenz von 4,6,8(14)-Trien-3-on-Steroiden. Fresenius Z Anal Chem 311:370–371
Schröder F (1984) Fluorescent sterols: Probe molecules of membrane structure and function. Prog Lipid Res 23: 97–113
Schulte KE, Rücker G, Fachmann H (1968) Ergosta 4,6,8(14), 22-Tetraenon-(3) als Inhaltstoff des Lärchenschwammes. Tetrahedron Lett 46:4763–4764
Westphal U (1978) Bindung von Progesteron und anderen Steroidhormonen and Proteine des Blutserums. Hoppe-Seyler's Z Physiol Chem 359: 431–447
White JD, Taylor SI (1970) Biosynthesis of ergosta-4,6,8(14), 22-tetraene-3-one. In vivo incorporation of a 1,4-dioxide. J Am Chem Soc 92:5811–5813
Author information
Authors and Affiliations
Additional information
Dedicated to Prof. Dr. F.-W. Zilliken on the occasion of his 65th birthday
This work was presented in part at the Annual Meeting of the ‘Gesellschaft für Biologische Chemie’, September 26–29, 1983, in Göttingen. For an abstract see: Hoppe-Seyler's Z. Physiol. Chem. (1983) 364: 1151–1152
Rights and permissions
About this article
Cite this article
Kempfle, M., Müller, R., Palluk, R. et al. Fluorescence of 3-keto-steroids in aqueous solution. Eur Biophys J 14, 29–35 (1986). https://doi.org/10.1007/BF00260400
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00260400