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Luminescence Resonance Energy Transfer to Study Conformational Changes in Membrane Proteins Expressed in Mammalian Cells
Published on: September 16, 2014
Effects of diffusion on energy transfer in solution using a microsecond decay time rhenium metal-ligand complex as
Józef Kuśba1, Grzegorz Piszczek2, Ignacy Gryczynski2
1Faculty of Applied Physics and Mathematics, Technical University of Gdansk, Gdansk, Poland.
Abstract:
We used resonance energy transfer and frequency-domain fluorometry to measure slow donor to acceptor diffusion in viscous media. The frequency-domain RET data were analyzed using a new numerical algorithm for predicting the donor intensity decay in the presence of diffusion occurring within the donor decay time. By the use of a rhenium metal-ligand complex as a microsecond decay time donor we were able to measure mutual donor-to-acceptor diffusion coefficients as low as 2 × 10-8 cm2/s. The availability of microsecond decay time luminophores and appropriate theory suggests the use of diffusion-enhanced energy transfer for measurement of diffusive processes and structural dynamics in biological systems.
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