Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/98127
Title: Weak glycolipid binding of a microdomain-tracer peptide correlates with aggregation and slow diffusion on cell membranes
Authors: Wohland, Thorsten
Kraut, Rachel
Geifman-Shochat, Susana
Lauterbach, Tim
Manna, Manoj
Ruhnow, Maria
Wisantoso, Yudi
Wang, Yaofeng
Matysik, Artur
Oglęcka, Kamila
Mu, Yuguang
Keywords: DRNTU::Science::Biological sciences
Issue Date: 2012
Source: Lauterbach, T., Manna, M., Ruhnow, M., Wisantoso, Y., Wang, Y., Matysik, A., et al. (2012). Weak Glycolipid Binding of a Microdomain-Tracer Peptide Correlates with Aggregation and Slow Diffusion on Cell Membranes. PLoS ONE, 7(12), e51222.
Series/Report no.: PLoS ONE
Abstract: Organized assembly or aggregation of sphingolipid-binding ligands, such as certain toxins and pathogens, has been suggested to increase binding affinity of the ligand to the cell membrane and cause membrane reorganization or distortion. Here we show that the diffusion behavior of the fluorescently tagged sphingolipid-interacting peptide probe SBD (Sphingolipid Binding Domain) is altered by modifications in the construction of the peptide sequence that both result in a reduction in binding to ganglioside-containing supported lipid membranes, and at the same time increase aggregation on the cell plasma membrane, but that do not change relative amounts of secondary structural features. We tested the effects of modifying the overall charge and construction of the SBD probe on its binding and diffusion behavior, by Surface Plasmon Resonance (SPR; Biacore) analysis on lipid surfaces, and by Fluorescence Correlation Spectroscopy (FCS) on live cells, respectively. SBD binds preferentially to membranes containing the highly sialylated gangliosides GT1b and GD1a. However, simple charge interactions of the peptide with the negative ganglioside do not appear to be a critical determinant of binding. Rather, an aggregation-suppressing amino acid composition and linker between the fluorophore and the peptide are required for optimum binding of the SBD to ganglioside-containing supported lipid bilayer surfaces, as well as for interaction with the membrane. Interestingly, the strength of interactions with ganglioside-containing artificial membranes is mirrored in the diffusion behavior by FCS on cell membranes, with stronger binders displaying similar characteristic diffusion profiles. Our findings indicate that for aggregation-prone peptides, aggregation occurs upon contact with the cell membrane, and rather than giving a stronger interaction with the membrane, aggregation is accompanied by weaker binding and complex diffusion profiles indicative of heterogeneous diffusion behavior in the probe population.
URI: https://hdl.handle.net/10356/98127
http://hdl.handle.net/10220/10899
ISSN: 1932-6203
DOI: http://dx.doi.org/10.1371/journal.pone.0051222
Rights: © 2012 The Authors. This paper was published in PLoS ONE and is made available as an electronic reprint (preprint) with permission of Lauterbach et al. The paper can be found at the following official DOI: [http://dx.doi.org/10.1371/journal.pone.0051222]. One print or electronic copy may be made for personal use only. Systematic or multiple reproduction, distribution to multiple locations via electronic or other means, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper is prohibited and is subject to penalties under law.
metadata.item.grantfulltext: open
metadata.item.fulltext: With Fulltext
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