Autonomous buckling of micrometer-sized lipid-protein membrane patches constructed by Dictyostelium discoideum

Journal of Biological Engineering

Taro Toyota¹  Kei Takahashi² 

[1] Research Center for Complex Systems Biology, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo 153-8902, Japan;Department of Basic Science, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo 153-8902, Japan

关键词: Dictyostelium discoideum;    Phosphatidylinositides;    Phosphatidylcholine;    Spin coating;    Substrate-supported lipid membrane;   

Others  :  1133468
DOI  :  10.1186/1754-1611-9-3

 received in 2014-10-31, accepted in 2015-01-11,  发布年份 2015

【 摘 要 】

Background

The cytosol of amoeba cells controls the membrane deformation during their motion in vivo. To investigate such ability of the cytosol of amoeba cell, Dictyostelium discoideum (Dictyostelium), in vitro, we used lipids extracted from Dictyostelium and commercially available phospholipids, and prepared substrate-supported lipid membrane patches on the micrometer scale by spin coating.

Results

We found that the spin coater holder, which has pores (pore size = 3.1 mm) of negative pressure to hold the cover glass induced the concave surface of the cover glass. The membrane lipid patches were formed at each position in the vicinity of the holder pores and their sizes were in the range of 2.7 to 3.2 × 10⁴ μm². After addition of the cytosol extracted from Dictyostelium to the lipid membrane patches, through time-lapse observation with a confocal laser scanning fluorescence microscope, we observed an autonomous buckling of the Dictyostelium lipid patches and localized behaviours of proteins found within.

Conclusion

The current method serves as the novel technique for the preparation of film patches in which the positions of patches are controlled by the holder pores without fabricating, modifying, and arranging the chemical properties of the solution components of lipids. The findings imply that lipid-binding proteins in the cytosol were adsorbed and accumulated within the Dictyostelium lipid patches, inducing the transformation of the cell-sized patch.

【 授权许可】

   
2015 Takahashi and Toyota; licensee BioMed Central.

【 图 表 】

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