Theory of Measurement of Forster-Type Energy Transfer in Macromolecules

We describe the theoretical basis of an unconventional method for the determination of the amount of energy transferred between two fluorophores by the Forster mechanism. The method involves an internal comparison made by separation of the fluorophores in situ (i.e., in the optical cell), for exampl...

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Bibliographic Details
Published inProceedings of the National Academy of Sciences - PNAS Vol. 80; no. 9; pp. 2579 - 2583
Main Authors Epe, Bernd, Steinhauser, Klaus G., Woolley, Paul
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences of the United States of America 01.05.1983
National Acad Sciences
Subjects
Anisotropy
Biological Sciences: Biophysics
Coefficients
Dyes
Emission spectra
Emissions intensity
Energy transfer
Macromolecules
Product labeling
Transfer RNA
Wavelengths
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Summary:We describe the theoretical basis of an unconventional method for the determination of the amount of energy transferred between two fluorophores by the Forster mechanism. The method involves an internal comparison made by separation of the fluorophores in situ (i.e., in the optical cell), for example, by means of enzymic digestion; it eliminates several important sources of error and it simplifies calculation while making maximal use of the information contained in the fluorescence spectra. The validity of the method is demonstrated by determination of the known distance between two modifiable sites on the transfer RNA molecule, and its usefulness is exemplified by its application to triangulation of the ribosome of Escherichia coli.
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ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.80.9.2579