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Vesicle fluctuation analysis of the effects of sterols on membrane bending rigidity

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Abstract

Sterols are regulators of both biological function and structure. The role of cholesterol in promoting the structural and mechanical stability of membranes is widely recognized. Knowledge of how the related sterols, lanosterol and ergosterol, affect membrane mechanical properties is sparse. This paper presents a comprehensive comparison of the effects of cholesterol, lanosterol, and ergosterol upon the bending elastic properties of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine giant unilamellar vesicles. Measurements are made using vesicle fluctuation analysis, a nonintrusive technique that we have recently improved for determining membrane bending rigidity. Giving a detailed account of the vesicle fluctuation analysis technique, we describe how the gravitational stabilization of the vesicles enhances image contrast, vesicle yield, and the quality of data. Implications of gravity on vesicle behaviour are also discussed. These recent modifications render vesicle fluctuation analysis an efficient and accurate method for determining how cholesterol, lanosterol, and ergosterol increase membrane bending rigidity.

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Notes

  1. The optical resolution of a membrane displacement is of the order 100 nm at a 100-fold magnification level.

  2. R1 and R2 are the radii of curvature in the principal directions obtained from the Weingarten matrix.

  3. The notation Σ lm will be used as an abbreviation of \( {\sum\nolimits_{l = 2}^{l_{{{\text{cut}}}} } {{\sum\nolimits_{m = - 1}^l {} }} } \) where lcut is the upper mode cutoff.

  4. The polar representation does not allow for overhangs, which results in the rejection of some contours. At this level it is necessary to make sure that all contours have the same direction of rotation in their polar representation.

Abbreviations

DMPC:

Dimyristoylphosphocholine

POPC:

1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine

VFA:

Vesicle fluctuation analysis

GUV:

Giant unilamellar vesicle

TLC:

Thin layer chromatography

EPR:

Electron paramagnetic resonance

NMR:

Nuclear magnetic resonance

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Acknowledgements

Thanks to Martin J. Zuckermann, Jenifer L. Thewalt and Till Boecking for helpful sterol advice. MEMPHYS-Center for Biomembrane Physics is supported by the Danish National Research Foundation. A.C.R. is a Julie Payette Scholar sponsored by the Natural Sciences and Engineering Research Council (NSERC) of Canada.

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Authors and Affiliations

  1. MEMPHYS Centre for Biomembrane Physics, Department of Chemistry, Technical University of Denmark, 2800 , Lyngby, Denmark

    Jonas Henriksen

  2. Department of Physics, University of Southern Denmark, Campusvej 55, 5239 , Odense, Denmark

    Amy C. Rowat

  3. Department of Chemistry, University of Southern Denmark, Campusvej 55, 5239 , Odense, Denmark

    John H. Ipsen

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  1. Jonas Henriksen
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  2. Amy C. Rowat
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  3. John H. Ipsen
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Correspondence to John H. Ipsen.

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Henriksen, J., Rowat, A.C. & Ipsen, J.H. Vesicle fluctuation analysis of the effects of sterols on membrane bending rigidity. Eur Biophys J 33, 732–741 (2004). https://doi.org/10.1007/s00249-004-0420-5

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  • DOI: https://doi.org/10.1007/s00249-004-0420-5

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