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The effect of the lipid-binding site of the ankyrin-binding domain of erythroid β-spectrin on the properties of natural membranes and skeletal structures

Cellular & Molecular Biology Letters volume 15, pages 406–423 (2010)Cite this article

Abstract

It was previously shown that the beta-spectrin ankyrin-binding domain binds lipid domains rich in PE in an ankyrin-dependent manner, and that its N-terminal sequence is crucial in interactions with phospholipids. In this study, the effect of the full-length ankyrin-binding domain of β-spectrin on natural erythrocyte and HeLa cell membranes was tested. It was found that, when encapsulated in resealed erythrocyte ghosts, the protein representing the full-length ankyrin-binding domain strongly affected the shape and barrier properties of the erythrocyte membrane, and induced partial spectrin release from the membrane, while truncated mutants had no effect. As found previously (Bok et al. Cell Biol. Int. 31 (2007) 1482–94), overexpression of the full-length GFP-tagged ankyrin-binding domain aggregated and induced aggregation of endogenous spectrin, but this was not the case with overexpression of proteins truncated at their N-terminus. Here, we show that the aggregation of spectrin was accompanied by the aggregation of integral membrane proteins that are known to be connected to spectrin via ankyrin, i.e. Na+K+ATP-ase, IP3 receptor protein and L1 CAM. By contrast, the morphology of the actin cytoskeleton remained unchanged and aggregation of cadherin E or N did not occur upon the overexpression of either full-length or truncated ankyrin-binding domain proteins. The obtained results indicate a substantial role of the lipid-binding part of the β-spectrin ankyrin-binding domain in the determination of the membrane and spectrin-based skeleton functional properties.

Abbreviations

DiD:

1,1’dioctadecyl 3,3,3’3’tetramethylindocarbocyanine

PC:

phosphatidylcholine

PE:

phosphatidylethanolamine

SDS PAGE:

SDS polyacrylamide gel electrophoresis

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Authors and Affiliations

  1. Laboratory of Cytobiochemistry, Faculty of Biotechnology, University of Wroclaw, Przybyszewskiego 63/77, 51-148, Wrocław, Poland

    Anna Chorzalska, Agnieszka Łach, Marcin Wolny, Anita Hryniewicz-Jankowska, Adam Kolondra & Aleksander F. Sikorski

  2. Wroclaw University of Technology, Wyb. Wyspiańskiego 27, 50-370, Wrocław, Poland

    Tomasz Borowik & Marek Langner

  3. Academic Centre for Biotechnology of Lipid Aggregates, Przybyszewskiego 63/77, 51-148, Wrocław, Poland

    Marek Langner & Aleksander F. Sikorski

Authors
  1. Anna Chorzalska
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  2. Agnieszka Łach
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  3. Tomasz Borowik
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  4. Marcin Wolny
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  5. Anita Hryniewicz-Jankowska
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  6. Adam Kolondra
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  7. Marek Langner
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  8. Aleksander F. Sikorski
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Correspondence to Aleksander F. Sikorski.

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Chorzalska, A., Łach, A., Borowik, T. et al. The effect of the lipid-binding site of the ankyrin-binding domain of erythroid β-spectrin on the properties of natural membranes and skeletal structures. Cell Mol Biol Lett 15, 406–423 (2010). https://doi.org/10.2478/s11658-010-0012-6

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  • DOI: https://doi.org/10.2478/s11658-010-0012-6

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Cellular & Molecular Biology Letters

ISSN: 1689-1392

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