Fluorescence Energy Transfer as a Structural Probe in Membranes and Membrane-Bound Proteins
In 1972 Radda and Vanderkooi (Radda 72) wrote “Fluorescence has been used in biochemical studies for many years, yet it is only fairly recently that its potential to the study of problems associated with membrane-related phenomena has been realized,” and in 1976 Badley (Badley 76) added “... it is only recently that fluorescence methods have been used in a more problem-oriented way in contrast to the, perhaps inevitable, initial rush of data primarily concerned with establishing the method as one of ‘high potential’.” I might begin by noting that as of 1982 only a handful of fluorescence experiments involving membranes have made full and rigorous use of the physical phenomena accessible to the fluorescence spectroscopist: quantum yield, spectral shifts, emission anisotropy, and resonance energy transfer including their multiple interactions. The first three of these phenomena have been treated in other lectures of this Advanced Study Institute. I will concentrate on energy transfer, but, as we shall see, it is quite useless and even misleading to try to analyze it without considering quantum yield and anisotropy, both in the steady state and time-resolved domains. Therefore, in the first few sections I will lay the groundwork necessary for the analysis of resonance energy transfer experiments.
KeywordsEnergy Transfer Resonance Energy Transfer Transition Moment Orientation Factor Energy Transfer Efficiency
Unable to display preview. Download preview PDF.
- Antonov-Romanovsky, V. V., and Galanin, M. D. (1957) Theoretical derivation of the law of luminescence decay for the case of resonance quenching. Optika i Spectroscopia 3:389–391.Google Scholar
- Badley, R. A., (1976) XXX in “Modern Fluorescence Spectroscopy,” (E. L. Whery, ed.), Plenum Press, New York, pp. 91–168.Google Scholar
- Eisinger, J., and Flores, J. (1983) The cytosol-membrane interface of human erythrocytes — a resonance energy transfer study. Biophysical J. (in press).Google Scholar
- FOrster, T. (1949) Experimentelle und theoretische untersuchung des zwischenmolekularen abergangs vom electronenanregungsenergie. Z. Naturforsch. 4a:322–327.Google Scholar
- Förster, T. (1951) “Fluoreszenz Organischer Verbindung,” Vandenhoeck and Ruprecht, Gottingen.Google Scholar
- Jones, R. E. (1970) Nanosecond fluorimetry. Ph.D. thesis, Stanford University.Google Scholar
- Perrin, F. (1926) Polarisation de la lumière de fluorescence. Vie moyenne des molécules dans l’etat excité. J. Phys. (Paris) 7:390–401.Google Scholar
- Radda, G. K., and Vanderkooi, J. (1972) Can fluorescent probes tell us anything about membranes? Biochim. Biophys. Acta 265:509–549.Google Scholar
- Rozman, I. M. (1958) Theory of quenching of luminescence in solutions. Optika i Spectroscopiya 4:536–538.Google Scholar
- Soleillet, P. (1929) Sur les paramètres caractérisant la polarisation partielle de la lumière dans les phénomènes de fluorescence. Ann. Physique 12:23–97.Google Scholar