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Monitoring Live Cell Membrane Lipid Encounter Dynamics with DNA Probe
Author(s) -
You Mingxu
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.629.3
Subject(s) - lipid raft , microbiology and biotechnology , membrane , membrane fluidity , cell membrane , cell , membrane lipids , cell signaling , biology , peripheral membrane protein , chemistry , biophysics , signal transduction , membrane protein , biochemistry , integral membrane protein
Lipids are ubiquitous membrane molecules that broadly involved in regulating signal transduction. Instead of homogeneously distributed, several lipid molecules clustered to form lipid rafts on cell membranes. These lipid rafts separate cell membranes into small pre‐organized compartmentalization. Abnormal level and interaction of lipids has been implicated in the pathogenesis of cancers, lysosomal storage disorders, peripheral neuropathies and secretory diarrhea. The disruption of lipid‐lipid and lipid‐protein interactions is important for pathogenesis of these lipid‐involved diseases. However, studying transient lipid encounter kinetics, especially on live cell membrane, remains a technical challenge. We have developed a DNA‐based probe to monitor lipid encounter kinetics on live leukemia cell membrane. A specific DNA toehold‐mediated strand displacement reaction transduces transient membrane encounter events into readable cumulative fluorescence signals. Our data indicates that previously untraceable cell membrane interaction kinetics among model diacyllipids, cholesterols and tocopherols can now be monitored with standard fluorescence microscopy and flow cytometry. We believe this DNA probe can be potentially used to study lipid encounter rates and preferences during various cell membrane signaling events.