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Originally published as Biophys J. BioFAST on March 24, 2006.
doi:10.1529/biophysj.105.070839
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Biophysical Journal 90:4452-4465 (2006)
© 2006 The Biophysical Society

Coexisting Domains in the Plasma Membranes of Live Cells Characterized by Spin-Label ESR Spectroscopy

Musti J. Swamy, Laura Ciani, Mingtao Ge, Andrew K. Smith, David Holowka, Barbara Baird and Jack H. Freed

Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14853

Correspondence: Address reprint requests to Jack H. Freed, Tel.: 607-255-3647; Fax: 607-255-6969; E-mail: jhf{at}ccmr.cornell.edu.

The importance of membrane-based compartmentalization in eukaryotic cell function has become broadly appreciated, and a number of studies indicate that these eukaryotic cell membranes contain coexisting liquid-ordered (Lo) and liquid-disordered (Ld) lipid domains. However, the current evidence for such phase separation is indirect, and so far there has been no direct demonstration of differences in the ordering and dynamics for the lipids in these two types of regions or their relative amounts in the plasma membranes of live cells. In this study, we provide direct evidence for the presence of two different types of lipid populations in the plasma membranes of live cells from four different cell lines by electron spin resonance. Analysis of the electron spin resonance spectra recorded over a range of temperatures, from 5 to 37°C, shows that the spin-labeled phospholipids incorporated experience two types of environments, Lo and Ld, with distinct order parameters and rotational diffusion coefficients but with some differences among the four cell lines. These results suggest that coexistence of lipid domains that differ significantly in their dynamic order in the plasma membrane is a general phenomenon. The Lo region is found to be a major component in contrast to a model in which small liquid-ordered lipid rafts exist in a ‘sea’ of disordered lipids. The results on ordering and dynamics for the live cells are also compared with those from model membranes exhibiting coexisting Lo and Ld phases.




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