کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1959897 1057947 2006 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Lateral Mobility of Membrane-Binding Proteins in Living Cells Measured by Total Internal Reflection Fluorescence Correlation Spectroscopy
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Lateral Mobility of Membrane-Binding Proteins in Living Cells Measured by Total Internal Reflection Fluorescence Correlation Spectroscopy
چکیده انگلیسی

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.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: - Volume 91, Issue 9, 1 November 2006, Pages 3456–3464
نویسندگان
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