End-to-end diffusion coefficients and distance distributions from fluorescence energy transfer measurements: enhanced resolution by using multiple donors with different lifetimes

Ignacy Gryczynski; Joseph R. Lakowicz; Jozef Kusba

doi:10.1117/12.182763

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17 August 1994 End-to-end diffusion coefficients and distance distributions from fluorescence energy transfer measurements: enhanced resolution by using multiple donors with different lifetimes

Ignacy Gryczynski, Joseph R. Lakowicz, Jozef Kusba

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Proceedings Volume 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV; (1994) https://doi.org/10.1117/12.182763
Event: OE/LASE '94, 1994, Los Angeles, CA, United States

Abstract

We describe a method to improve the resolution of donor-to- acceptor distance distributions in molecules which are flexing on the timescale of the fluorescence lifetime. We measured the time- dependent donor decays of two donor (D)-acceptor (A) pairs, where the donor lifetimes were substantially different. The donors were an indole residue (5.7 ns) and a naphthalene residue (24.4 ns). The same dansyl acceptor was used for both D-A pairs. The donor decays are complex due to both a distribution of D-A distances and D-A diffusion. Using the donor decay data for each D-A pair alone, it is difficult to resolve both the distance distribution and the D-to-A diffusion coefficient. However, these values are unambiguously recovered from global analysis of the data from both D-A pairs. Simulations were also used to demonstrate the increased reduction of global analysis with different lifetime donors to obtain distance distribution parameters in the presence of D-A diffusion.

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Ignacy Gryczynski, Joseph R. Lakowicz, and Jozef Kusba "End-to-end diffusion coefficients and distance distributions from fluorescence energy transfer measurements: enhanced resolution by using multiple donors with different lifetimes", Proc. SPIE 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV, (17 August 1994); https://doi.org/10.1117/12.182763

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