Raft-like domain formation in large unilamellar vesicles probed by the fluorescent phospholipid analogue, C12NBD-PC

Biochimica Et Biophysica Acta
Virginie CosteMiglena I Angelova

Abstract

The liquid-ordered/disordered-phase domain co-existence in large unilamellar vesicle membranes consisting of phosphatidylcholine:sphingomyelin (2:1) with different amounts of cholesterol has been examined using a concentration-dependent self-quenching of a single reporter molecule, C12NBD-PC. A temperature-dependent decrease of fluorescence intensity was associated with the expected formation and increase of l(o)-phase membrane fraction in the vesicles. The result is consistent with exclusion of the fluorescent probe from the liquid-ordered phase which partitions preferentially into the liquid-disordered phase membrane domains. This leads to an increase of the local concentration of fluorophore in the liquid-disordered phase and a decrease of the quantum yield. This effect was used to obtain a quantitative estimation of the fraction of the vesicle membrane occupied by the liquid-ordered phase, Phi(o), as a function of temperature and cholesterol content between 0 and 45 mol%. The value of Phi(o) was related to the assumed partition coefficient k(p) of probe between liquid-ordered/disordered phases. For large unilamellar vesicles containing 20 and 4 mol% cholesterol and probe, respectively, with k(p) = 0 (probe completely exclud...Continue Reading

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Citations

Jul 23, 2008·Journal of the American Chemical Society·Adekunle I ElegbedeSanku Mallik
Jan 27, 2009·Biophysical Journal·James H DavisJanos Juhasz
Jan 9, 2010·Biochimica Et Biophysica Acta·M T Stöckl, A Herrmann
Dec 23, 2009·Biochimica Et Biophysica Acta·Daniel M Carter RamirezLinda J Johnston

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