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MBC in Press, published online ahead of print February 6, 2008
Mol. Biol. Cell 10.1091/mbc.E07-12-1208

A more recent version of this article appeared on April 1, 2008
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Submitted on December 6, 2007
Revised on January 10, 2008
Accepted on January 24, 2008

Diffusion Coefficient of Fluorescent Phosphatidylinositol 4,5-bisphosphate (PIP2) in the Plasma Membrane of Cells

Urszula Golebiewska, Marian Nyako, William Woturski, Irina Zaitseva, and Stuart McLaughlin

Department of Physiology and Biophysics, Health Science Center, Stony Brook University, Stony Brook, NY 11794-8661

Monitoring Editor: Tom U. Martin

Phosphatidylinositol 4,5-bisphosphate (PIP2) controls a surprisingly large number of processes in cells. Thus, a number of investigators have suggested that there might be different pools of PIP2 on the inner leaflet of the plasma membrane. If a significant fraction of PIP2 is bound electrostatically to unstructured clusters of basic residues on membrane proteins, the PIP2 diffusion constant, D, should be reduced. We microinjected micelles of Bodipy TMR-PIP2 into cells and measured D on the inner leaflet of fibroblasts and epithelial cells using fluorescence correlation spectroscopy (FCS). The average value from all cell types was D = 0.8 ± 0.2 µm2/s (n = 218, T = 25°C). This is threefold lower than the D in blebs formed on Rat1 cells, D = 2.5 ± 0.8 µm2/s (n = 26). It is also significantly lower than the D in the outer leaflet or in giant unilamellar vesicles and the diffusion coefficient for other lipids on the inner leaflet of these cell membranes. The simplest interpretation is that ~2/3 of the PIP2 on inner leaflet of these plasma membranes is bound reversibly.

Address correspondence to: Stuart McLaughlin (Stuart.McLaughlin{at}stonybrook.edu)




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