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Lateral Mobility of Membrane-Binding Proteins in Living Cells Measured by Total Internal Reflection Fluorescence Correlation Spectroscopy

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Title: Lateral Mobility of Membrane-Binding Proteins in Living Cells Measured by Total Internal Reflection Fluorescence Correlation Spectroscopy
Authors: Ohsugi, Yu Browse this author
Saito, Kenta Browse this author
Tamura, Mamoru Browse this author
Kinjo, Masataka Browse this author →KAKEN DB
Keywords: Fluorescence correlation spectroscopy
Total internal reflection fluorescence
Farnesylated EGFP
Plasma membrane
Diffusion constant
Single-molecule tracking
Issue Date: Nov-2006
Publisher: The Biophysical Society
Journal Title: Biophysical Journal
Volume: 91
Issue: 9
Start Page: 3456
End Page: 3464
Publisher DOI: 10.1529/biophysj.105.074625
Abstract: Total internal reflection fluorescence correlation spectroscopy (TIR-FCS) allows us to measure diffusion constants and the number of fluorescent molecules in a small area of an evanescent field generated on the objective of a microscope. The application of TIR-FCS makes possible the characterization of reversible association and dissociation rates between fluorescent ligands and their receptors in supported phospholipid bilayers. Here, for the first time, we extend TIR-FCS to a cellular application for measuring the lateral diffusion of a membrane-binding fluorescent protein, farnesylated EGFP, on the plasma membranes of cultured HeLa and COS7 cells. We detected two kinds of diffusional motion—fast three-dimensional diffusion (D1) and much slower two-dimensional diffusion (D2), simultaneously. Conventional FCS and single-molecule tracking confirmed that D1 was free diffusion of farnesylated EGFP close to the plasma membrane in cytosol and D2 was lateral diffusion in the plasma membrane. These results suggest that TIR-FCS is a powerful technique to monitor movement of membrane-localized molecules and membrane dynamics in living cells.
Type: article (author version)
URI: http://hdl.handle.net/2115/16923
Appears in Collections:電子科学研究所 (Research Institute for Electronic Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 金城 政孝

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