Global analysis of distance distribution data for donor-acceptor pairs with different Forster distances

Wieslaw M. Wiczk; Peggy S. Eis; Mayer N. Fishman; Michael L. Johnson; Joseph R. Lakowicz

doi:10.1117/12.17757

1 May 1990 Global analysis of distance distribution data for donor-acceptor pairs with different Förster distances

Wieslaw M. Wiczk, Peggy S. Eis, Mayer N. Fishman, Michael L. Johnson, Joseph R. Lakowicz

Proceedings Volume 1204, Time-Resolved Laser Spectroscopy in Biochemistry II; (1990) https://doi.org/10.1117/12.17757
Event: OE/LASE '90, 1990, Los Angeles, CA, United States

Abstract

Distance distributions of a flexible molecule were recovered from steady-state fluorescence energy transfer measurements with the method suggested by Cantor and Pechukas In this method, the Forster distance (R₀) is varied by attaching different donor-acceptor (D-A) pairs to the linker of interest. The distance distributions are recovered from the energy transfer efficiency measurements on D-A pairs with different R0s, followed by non-linear least squares fitting to distance distribution models. Thirteen compounds were synthesized with R0s ranging from 6-32 angstroms. Each compound contained a tryptamine donor linked by an alkyl chain (~10 carbon atoms) to one of thirteen different acceptors. The distance distributions were recovered by fitting the transfer efficiencies of the D-A pairs to both Gaussian and skewed Gaussian models. Our results indicate that the shape of the distribution is best described by a skewed Gaussian model. This is the first reported measurement of a skewed Gaussian distance distribution. In addition, the results of the Cantor and Pechuka's method and analysis were found to be consistent with distance distributions recovered from time-resolved frequency-domain fluorometry.

Citation Download Citation

Wieslaw M. Wiczk, Peggy S. Eis, Mayer N. Fishman, Michael L. Johnson, and Joseph R. Lakowicz "Global analysis of distance distribution data for donor-acceptor pairs with different Förster distances", Proc. SPIE 1204, Time-Resolved Laser Spectroscopy in Biochemistry II, (1 May 1990); https://doi.org/10.1117/12.17757

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