Lateral diffusion contributes to FRET from lanthanide-tagged membrane proteins
Tien-Hung Lan1, Guangyu Wu1, Nevin A Lambert1
1Department of Pharmacology and Toxicology, Medical College of Georgia, Georgia Regents University, Augusta, GA, USA.
Biochemical and Biophysical Research Communications
|June 30, 2015
Summary
Diffusion significantly boosts Förster resonance energy transfer (FRET) for membrane proteins using long-lived lanthanide donors. This effect complicates proximity measurements but enables real-time monitoring of membrane diffusion in living cells.
Area of Science:
- Biophysics
- Cell Biology
- Spectroscopy
Background:
- Förster resonance energy transfer (FRET) is a distance-dependent process used to study molecular interactions.
- Long-lived lanthanide donors offer advantages for time-resolved FRET but their long excited-state lifetimes may allow for diffusion effects.
- Understanding diffusion's impact is crucial for interpreting FRET measurements in dynamic biological systems.
Purpose of the Study:
- To investigate the impact of diffusion on Förster resonance energy transfer (FRET) between membrane proteins labeled with lanthanide donors.
- To determine if diffusion enhances FRET when donor and acceptor molecules are mobile.
- To explore the potential of diffusion-enhanced FRET for monitoring membrane protein dynamics.
Main Methods:
- Utilizing time-resolved Förster resonance energy transfer (FRET) measurements.
- Labeling membrane proteins with lanthanide donors and suitable acceptors.
- Analyzing FRET efficiency in relation to protein mobility and diffusion distances.
Main Results:
- Demonstrated that diffusion significantly enhances FRET between membrane proteins labeled with lanthanide donors.
- Observed that diffusion becomes a major factor when diffusion distances approach the Förster radius during the donor's long excited-state lifetime.
- Showcased that this diffusion-enhanced FRET can be observed in real-time in living cells.
Conclusions:
- Diffusion dramatically enhances Förster resonance energy transfer (FRET) for mobile membrane proteins labeled with long-lived lanthanide donors.
- This phenomenon presents challenges for interpreting FRET data regarding protein association but offers a novel method for real-time diffusion monitoring.
- Diffusion-enhanced FRET provides a new tool to study the dynamics and organization of membrane domains in living cells.
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