Fluorescence lifetime tuning--a novel approach to study flip-flop kinetics in supported phospholipid bilayers
Language English Country Netherlands Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Time Factors MeSH
- Chemistry Techniques, Analytical methods MeSH
- Diffusion MeSH
- Fluorescence * MeSH
- Fluorescent Dyes chemistry MeSH
- Phosphatidylcholines chemistry MeSH
- Phosphatidylethanolamines chemistry MeSH
- Phosphatidylserines chemistry MeSH
- Phospholipids chemistry MeSH
- Kinetics MeSH
- Lipid Bilayers chemistry MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 1,2-dielaidoylphosphatidylethanolamine MeSH Browser
- 1,2-dioleoylphosphatidylserine MeSH Browser
- 1,2-oleoylphosphatidylcholine MeSH Browser
- Fluorescent Dyes MeSH
- Phosphatidylcholines MeSH
- Phosphatidylethanolamines MeSH
- Phosphatidylserines MeSH
- Phospholipids MeSH
- Lipid Bilayers MeSH
In the present work we introduce a straightforward fluorescent assay that can be applied in studies of the transbilayer movement (flip-flop) of fluorescent lipid analogues across supported phospholipid bilayers (SPBs). The assay is based on the distance dependent fluorescence quenching by light absorbing surfaces. Applied to SPBs this effect leads to strong differences in fluorescence lifetimes when the dye moves from the outer lipid leaflet to the leaflet in contact with the support. Herein, we present the basic principles of this novel approach, and comment on its advantages over the commonly used methods for investigating flip-flop dynamics across lipid bilayers. We test the assay on the fluorescent lipid analog Atto633-DOPE and the 3-hydroxyflavone F2N12S probe in SPBs composed of DOPC/ DOPS lipids. Moreover, we compare and discuss the flip-flop rates of the probes with respect to their lateral diffusion coefficients.
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