Cell-derived GPMVs (giant plasma-membrane vesicles) enable investigation of lipid phase separation in a system with appropriate biological complexity under physiological conditions, and in the present study were used to investigate the cholesterol-dependence of domain formation and stability. The cholesterol level is directly related to the abundance of the liquid-ordered phase fraction, which is the majority phase in vesicles from untreated cells. Miscibility transition temperature depends on cholesterol and correlates strongly with the presence of detergent-insoluble membrane in cell lysates. Fluorescence correlation spectroscopy reveals two distinct diffusing populations in phase-separated cell membrane-derived vesicles whose diffusivities correspond well to diffusivities in both model systems and live cells. The results of the present study extend previous observations in purified lipid systems to the complex environment of the plasma membrane and provide insight into the effect of cholesterol on lipid phase separation and abundance.
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November 11 2009
Cholesterol-dependent phase separation in cell-derived giant plasma-membrane vesicles Available to Purchase
Ilya Levental;
Ilya Levental
*Department of Bioengineering, University of Pennsylvania, 1010 Vagelos Laboratories, 3340 Smith Walk, Philadelphia, PA 19104, U.S.A.
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Fitzroy J. Byfield;
Fitzroy J. Byfield
†Institute for Medicine and Engineering, University of Pennsylvania, 1010 Vagelos Laboratories, 3340 Smith Walk, Philadelphia, PA 19104, U.S.A.
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Pramit Chowdhury;
Pramit Chowdhury
‡Department of Chemistry, University of Pennsylvania, 1010 Vagelos Laboratories, 3340 Smith Walk, Philadelphia, PA 19104, U.S.A.
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Feng Gai;
Feng Gai
‡Department of Chemistry, University of Pennsylvania, 1010 Vagelos Laboratories, 3340 Smith Walk, Philadelphia, PA 19104, U.S.A.
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Tobias Baumgart;
Tobias Baumgart
‡Department of Chemistry, University of Pennsylvania, 1010 Vagelos Laboratories, 3340 Smith Walk, Philadelphia, PA 19104, U.S.A.
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Paul A. Janmey
Paul A. Janmey
1
§Department of Physics, University of Pennsylvania, 1010 Vagelos Laboratories, 3340 Smith Walk, Philadelphia, PA 19104, U.S.A.
∥Department of Physiology, University of Pennsylvania, 1010 Vagelos Laboratories, 3340 Smith Walk, Philadelphia, PA 19104, U.S.A.
1To whom correspondence should be addressed (email [email protected]).
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Publisher: Portland Press Ltd
Received:
August 17 2009
Revision Received:
September 28 2009
Accepted:
October 07 2009
Accepted Manuscript online:
October 07 2009
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2009 Biochemical Society
2009
Biochem J (2009) 424 (2): 163–167.
Article history
Received:
August 17 2009
Revision Received:
September 28 2009
Accepted:
October 07 2009
Accepted Manuscript online:
October 07 2009
Connected Content
A commentary has been published:
Biomembrane liquid–liquid phase separation and detergent resistance: a relationship strengthened
Citation
Ilya Levental, Fitzroy J. Byfield, Pramit Chowdhury, Feng Gai, Tobias Baumgart, Paul A. Janmey; Cholesterol-dependent phase separation in cell-derived giant plasma-membrane vesicles. Biochem J 1 December 2009; 424 (2): 163–167. doi: https://doi.org/10.1042/BJ20091283
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